Research activity information

• Metabolism of testosterone and progesterone by cytochrome P450 2C19 allelic variants

  Shiori Takeji, Mai Okada, Shu Hayashi, Kengo Kanamaru, Yuichi Uno, Hiromasa Imaishi, Tomohide Uno

  Abstract CYP2C19 is a member of the human microsomal cytochrome P450 (CYP). Significant variation in CYP2C19 levels and activity can be attributed to polymorphisms in this gene. Wildtype CYP2C19 and 13 mutants (CYP2C19.1B, CYP2C19.5A, CYP2C19.5B, CYP2C19.6, CYP2C19.8, CYP2C19.9, CYP2C19.10, CYP2C19.11, CYP2C19.13, CYP2C19.16, CYP2C19.19, CYP2C19.23, CYP2C19.30, and CYP2C19.33) were coexpressed with NADPH‐cytochrome P450 reductase in Escherichia coli. Hydroxylase activity toward testosterone and progesterone was also examined. Ten CYP2C19 variants showed Soret peaks (450 nm) typical of P450 in the reduced CO‐difference spectra. CYP2C19.11 and CYP2C19.23 showed higher testosterone 11α, 16α‐/17‐ and progesterone 6β‐,21‐,16α‐/17α‐hydroxylase activities than CYP2C19.1B. CYP2C19.6, CYP2C19.16, CYP2C19.19, and CYP2C19.30 showed lower activity than CYP2C19.1B. CYP2C19.9, CYP2C19.10. CYP2C19.13, and CYP2C19.33 showed different hydroxylation activities than CYP2C19.1B. These results indicated that CYP2C19 variants have very different substrate specificities for testosterone and progesterone.

  Wiley, Oct. 2023, Biopharmaceutics & Drug Disposition, English

  [Refereed]

  Scientific journal


• Locations of Rab, Allatotropin, Prothoracicotropic hormone and Period in the larval brain, corpus allatum and frontal ganglion of Bombyx mori

  Asuka MATSUI, Makoto TOKUSHIGE, Akira MIZOGUCHI, Kengo KANAMARU, Katsuhiko SAKAMOTO, Yuichi UNO, Tomohide UNO

  Biology Centre, AS CR, Mar. 2023, European Journal of Entomology, 120, 93 - 104, English

  [Refereed]

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Argon-ion-induced mutations in Arabidopsis EGY1 gene affect chloroplast development in leaf guard cells.

  Sanjaya A, Muramatsu R, Sato S, Suzuki M, Sasaki S, Ishikawa H, Fujii Y, Asano M, Itoh R, Kanamaru K, Ohbu S, Abe T, Kazama Y, Fujiwara M

  Dec. 2022, RIKEN Accelerator Progress Report, English

  [Refereed]

  Research institution


• Localization of SNARE proteins in the brain and corpus allatum of Bombyx mori

  Mako Sasao, Tomohide Uno, Risa Kitagawa, Asuka Matsui, Fumika Toryu, Akira Mizoguchi, Kengo Kanamaru, Katsuhiko Sakamoto, Yuichi Uno

  Springer Science and Business Media LLC, Sep. 2022, Histochemistry and Cell Biology, 159(2) (2), 199 - 208, English

  [Refereed]

  Scientific journal


• Arabidopsis EGY1 Is Critical for Chloroplast Development in Leaf Epidermal Guard Cells

  Alvin Sanjaya, Ryohsuke Muramatsu, Shiho Sato, Mao Suzuki, Shun Sasaki, Hiroki Ishikawa, Yuki Fujii, Makoto Asano, Ryuuichi D. Itoh, Kengo Kanamaru, Sumie Ohbu, Tomoko Abe, Yusuke Kazama, Makoto T. Fujiwara

  In Arabidopsis thaliana, the Ethylene-dependent Gravitropism-deficient and Yellow-green 1 (EGY1) gene encodes a thylakoid membrane-localized protease involved in chloroplast development in leaf mesophyll cells. Recently, EGY1 was also found to be crucial for the maintenance of grana in mesophyll chloroplasts. To further explore the function of EGY1 in leaf tissues, we examined the phenotype of chloroplasts in the leaf epidermal guard cells and pavement cells of two 40Ar17+ irradiation-derived mutants, Ar50-33-pg1 and egy1-4. Fluorescence microscopy revealed that fully expanded leaves of both egy1 mutants showed severe chlorophyll deficiency in both epidermal cell types. Guard cells in the egy1 mutant exhibited permanent defects in chloroplast formation during leaf expansion. Labeling of plastids with CaMV35S or Protodermal Factor1 (PDF1) promoter-driven stroma-targeted fluorescent proteins revealed that egy1 guard cells contained the normal number of plastids, but with moderately reduced size, compared with wild-type guard cells. Transmission electron microscopy further revealed that the development of thylakoids was impaired in the plastids of egy1 mutant guard mother cells, guard cells, and pavement cells. Collectively, these observations demonstrate that EGY1 is involved in chloroplast formation in the leaf epidermis and is particularly critical for chloroplast differentiation in guard cells.

  MDPI AG, Jun. 2021, Plants, 10(6) (6), 1254 - 1254

  [Refereed]

  Scientific journal


• An Argon-Ion-Induced Pale Green Mutant of Arabidopsis Exhibiting Rapid Disassembly of Mesophyll Chloroplast Grana.

  Alvin Sanjaya, Yusuke Kazama, Kotaro Ishii, Ryohsuke Muramatsu, Kengo Kanamaru, Sumie Ohbu, Tomoko Abe, Makoto T Fujiwara

  Argon-ion beam is an effective mutagen capable of inducing a variety of mutation types. In this study, an argon ion-induced pale green mutant of Arabidopsis thaliana was isolated and characterized. The mutant, designated Ar50-33-pg1, exhibited moderate defects of growth and greening and exhibited rapid chlorosis in photosynthetic tissues. Fluorescence microscopy confirmed that mesophyll chloroplasts underwent substantial shrinkage during the chlorotic process. Genetic and whole-genome resequencing analyses revealed that Ar50-33-pg1 contained a large 940 kb deletion in chromosome V that encompassed more than 100 annotated genes, including 41 protein-coding genes such as TYRAAt1/TyrA1, EGY1, and MBD12. One of the deleted genes, EGY1, for a thylakoid membrane-localized metalloprotease, was the major contributory gene responsible for the pale mutant phenotype. Both an egy1 mutant and F1 progeny of an Ar50-33-pg1 × egy1 cross-exhibited chlorotic phenotypes similar to those of Ar50-33-pg1. Furthermore, ultrastructural analysis of mesophyll cells revealed that Ar50-33-pg1 and egy1 initially developed wild type-like chloroplasts, but these were rapidly disassembled, resulting in thylakoid disorganization and fragmentation, as well as plastoglobule accumulation, as terminal phenotypes. Together, these data support the utility of heavy-ion mutagenesis for plant genetic analysis and highlight the importance of EGY1 in the structural maintenance of grana in mesophyll chloroplasts.

  Apr. 2021, Plants (Basel, Switzerland), 10(5) (5), English, International magazine

  [Refereed]

  Scientific journal


• Relationship between Rab and insulin-like proteins in the nervous system of Bombyx mori

  Tomohide Uno, Yusuke Ozakiya, Mako Sasao, Katsuhiko Sakamoto, Yasuo Yamauchi, Yuichi Uno, Kengo Kanamaru, Akira Mizoguchi

  Rab proteins are small GTP-binding proteins and are the largest family in the Ras GTPase superfamily and mediate vesicular transport in cells. Diverse insulin-like peptides, such as bombyxin, are synthesized in the brain and secreted into the haemolymph by the corpus allatum (CA). In the brain of Bombyx mori, Rabs are expressed in a specific area; however, which Rabs actually link the secretion of bombyxin remains unknown. A double-staining analysis of nine Rabs ( Rab1, 3, 6, 7, 14, 21, 26, 39 and X4) and bombyxin indicated that Rab3-, Rab7-, Rab39-and RabX4-immunohistochemical reactivity (ir) areas overlapped with bombyxin-ir in the brain and CA in B. mori, while Rab6-, Rab14-and Rab21-irs partially overlapped in the CA. Rab1-ir occurred in the other immunopositive areas in CA. Rab26-ir did not occur in the brain. Rab39-ir occurred in UNC104, Rab39- effector,-immunopositive neurons in the brain and CA. Thus, Rab3, 7, 39 and X4 may regulate the exocytosis of bombyxin.

  CZECH ACAD SCI, INST ENTOMOLOGY, 2021, EUROPEAN JOURNAL OF ENTOMOLOGY, 118, 307 - 314, English

  [Refereed]

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• 5-Aminolevulinic acid fermentation using engineered Saccharomyces cerevisiae

  Kiyotaka Y. Hara, Masaru Saito, Hiroko Kato, Kana Morikawa, Hiroshi Kikukawa, Hironari Nomura, Takanori Fujimoto, Yoko Hirono-Hara, Shigeyuki Watanabe, Kengo Kanamaru, Akihiko Kondo

  Springer Science and Business Media LLC, Dec. 2019, Microbial Cell Factories, 18(1) (1)

  [Refereed]

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Functional characterization of insect-specific RabX6 of Bombyx mori

  Tomohide UNO, Yusuke OZAKIYA, Masayuki FURUTANI, Katsuhiko SAKAMOTO, Yuichi UNO, Kengo KANAMARU, Akira MIZOGUCHI

  Jan. 2019, Histochemistry and Cell Biology, 151, 187 - 198, English

  [Refereed]

  Scientific journal


• Metabolism of steroids by cytochrome P450 2C9 variants.

  Tomohide Uno, Ryosuke Nakano, Rie Kitagawa, Mai Okada, kengo Kanamaru, Shinji Takenaka, Yuichi Uno, Hiromasa Imaishi

  CYP2C9 is a human microsomal cytochrome P450c (CYP). Much variation in CYP2C9 levels and activity can be attributed to polymorphisms of this gene. Wild-type CYP2C9 and ten mutants were co-expressed with NADPH-cytochrome P450 reductase in Escherichia coli. The hydroxylase activities toward steroids were examined. CYP2C9.2, CYP2C9.3, CYP2C9.4, CYP2C9.16, CYP2C9.28, CYP2C9.48 and

  Aug. 2018, Biopharmaceutics & Drug Disposition, 39(8) (8), 371 - 377, English

  [Refereed]

  Scientific journal


• Metabolism of 7-ethoxycoumarin, flavanone and steroids by cytochrome P450 2C9 variants

  Tomohide Uno, Ryosuke Nakano, Kengo Kanamaru, Shinji Takenaka, Yuichi Uno, Hiromasa Imaishi

  CYP2C9 is a human microsomal cytochrome P450c (CYP). Much of the variation in CYP2C9 levels and activity can be attributed to polymorphisms of this gene. Wild-type CYP2C9 and mutants were coexpressed with NADPH-cytochrome P450 reductase in Escherichia coli. The hydroxylase activities toward 7-ethoxycoumarin, flavanone and steroids were examined. Six CYP2C9 variants showed Soret peaks (450 nm) typical of P450 in reduced CO-difference spectra. CYP2C9.38 had the highest 7-ethoxycoumarin de-ethylase activity. All the CYP2C9 variants showed lower flavanone 6-hydroxylation activities than CYP2C9.1 (the wild-type). CYP2C9.38 showed higher activities in testosterone 6β-hydroxylation, progesterone 6β−/16α-hydroxylation, estrone 11α-hydroxylation and estradiol 6α-hydroxylation than CYP2C9.1. CYP2C9.40 showed higher testosterone 17-oxidase activity than CYP2C9.1 CYP2C9.8 showed higher estrone 16α-hydroxylase activity and CYP2C9.12 showed higher estrone 11α-hydroxylase activity. CYP2C9.9 and CYP2C9.10 showed similar activities to CYP2C9.1. These results indicate that the substrate specificity of CYP2C9.9 and CYP2C9.10 was not changed, but CYP2C9.8, CYP2C9.12 and CYP2C9.40 showed different substrate specificity toward steroids compared with CYP2C9.1 and especially CYP2C9.38 displayed diverse substrate specificities towards 7-ethoxycoumarin and steroids.

  John Wiley and Sons Ltd, Nov. 2017, Biopharmaceutics and Drug Disposition, 38(8) (8), 486 - 493, English

  [Refereed]

  Scientific journal


• Localization and functional analysis of the insect-specific RabX4 in the brain of Bombyx mori

  Tomohide Uno, Masayuki Furutani, Katsuhiko Sakamoto, Yuichi Uno, Kengo Kanamaru, Akira Mizoguchi, Susumu Hiragaki, Makio Takeda

  Rab proteins are small monomeric GTPases/GTP-binding proteins, which form the largest branch of the Ras superfamily. The different Rab GTPases are localized to the cytosolic face of specific intracellular membranes, where they function as regulators of distinct steps in membrane trafficking. RabX4 is an insect-specific Rab protein that has no close homolog in vertebrates. There is little information about insect-specific Rab proteins. RabX4 was expressed in Escherichia coli and subsequently purified. Antibodies against Bombyx mori RabX4 were produced in rabbits for western immunoblotting and immunohistochemistry. Western blotting of neural tissues revealed a single band, at approximately 26kD. RabX4-like immunohistochemical reactivity was restricted to neurons of the pars intercerebralis and dorsolateral protocerebrum in the brain. Further immunohistochemical analysis revealed that RabX4 colocalized with Rab6 and bombyxin in the corpus allatum, a neuronal organ that secretes neuropeptides synthesized in the brain into the hemolymph. RabX4 expression in the frontal ganglion, part of the insect stomatogastric nervous system that is found in most insect orders, was restricted to two neurons on the outer region and did not colocalize with allatotropin or Rab6. Furthermore, RNA interference of RabX4 decreased bombyxin expression levels in the brain. These findings suggest that RabX4 is involved in the neurosecretion of a secretory organ in Bombyx mori.

  WILEY, Sep. 2017, ARCHIVES OF INSECT BIOCHEMISTRY AND PHYSIOLOGY, 96(1) (1), English

  [Refereed]

  Scientific journal


• Rab proteins in the brain and corpus allatum of Bombyx mori

  Tomohide Uno, Masayuki Furutani, Chihiro Watanabe, Katsuhiko Sakamoto, Yuichi Uno, Kengo Kanamaru, Hiroshi Yamagata, Akira Mizoguchi, Makio Takeda

  In eukaryotic cells, Rab guanosine triphosphate-ases serve as key regulators of membrane-trafficking events, such as exocytosis and endocytosis. Rab3, Rab6, and Rab27 control the regulatory secretory pathway of neuropeptides and neurotransmitters. The cDNAs of Rab3, Rab6, and Rab27 from B. mori were inserted into a plasmid, transformed into Escherichia coli, and then subsequently purified. We then produced antibodies against Rab3, Rab6, and Rab27 of Bombyx mori in rabbits and rats for use in western immunoblotting and immunohistochemistry. Western immunoblotting of brain tissue revealed a single band at approximately 26 kDa. Immunohistochemistry results revealed that Rab3, Rab6, and Rab27 expression was restricted to neurons in the pars intercerebralis and dorsolateral protocerebrum of the brain. Rab3 and Rab6 co-localized with bombyxin, an insect neuropeptide. However, there was no Rab that co-localized with prothoracicotropic hormone. The corpus allatum secretes neuropeptides synthesized in the brain into the hemolymph. Results showed that Rab3 and Rab6 co-localized with bombyxin in the corpus allatum. These findings suggest that Rab3 and Rab6 are involved in neurosecretion in B. mori. This study is the first to report a possible relationship between Rab and neurosecretion in the insect corpus allatum.

  SPRINGER, Jul. 2016, HISTOCHEMISTRY AND CELL BIOLOGY, 146(1) (1), 59 - 69, English

  [Refereed]

  Scientific journal


• Natural variation in the glucose content of dilute sulfuric acid-pretreated rice straw liquid hydrolysates: implications for bioethanol production

  Takashi Goda, Hiroshi Teramura, Miki Suehiro, Kengo Kanamaru, Hideo Kawaguchi, Chiaki Ogino, Akihiko Kondo, Masanori Yamasaki

  Rice straw is a promising resource for bioethanol production. Because the glucose content of pretreatment liquid hydrolysates is highly correlated with ethanol yield, the selection of appropriate rice cultivars is essential. The glucose content in liquid hydrolysates of pretreated rice straws of 208 diverse cultivars was evaluated in natural field in 2013 and 2014 using a novel high-throughput system. The glucose content of the rice straw samples varied across cultivars and was affected by environmental factors such as temperature and solar radiation. Several high-quality cultivars exhibiting high glucose content in both years were identified. The results of this study can aid in development of novel rice cultivars suitable as both feedstocks for bioethanol production and cooking.

  TAYLOR & FRANCIS LTD, May 2016, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 80(5) (5), 863 - 869, English

  [Refereed]

  Scientific journal


• Functional characterization of CYP1A9 and CYP1C1 from Anguillus japonica

  Tomohide Uno, Chiho Izumi, Shinji Takenaka, Takeshi Yanase, Hiromasa Imaishi, Kengo Kanamaru, Hiroshi Yamagata, Yoshio Kaminishi, Takao Itakura

  We evaluated the metabolism of several herbicides and progesterone by two P450 proteins (CYP1A9 and CYP1C1) from Japanese eel (Anguilla japonica). Expression vectors harboring CYP1A9 and CYP1C1 sequences were introduced into Escherichia coli. E. coli membrane fractions were incubated with each substrate, and the metabolites were analyzed. CYP1A9 and CYP1C1 deethylated 7-ethoxycoumarin and phenacetin, and demethylated chlorotoluron, diuron, and linuron. CYP1C1 specifically hydroxlyated progesterone at the 6 beta and 16 alpha positions. Five amino acids of CYP1A9 related to substrate binding were selected for mutation analyses [CYP1A9(F128A), CYP1A9(F229A), CYP1A9(F263A), CYP1A9(V387A), and CYP1A9(I391A)]. Two variants, CYP1A9(F229A) and CYP1A9(F128A), changed the ratio of 16a hydroxyprogesterone to 613 hydroxyprogesterone. Among all the variants, CYP1A9(F263A) showed the highest activity towards substrates used. CYP1A9(V387A) and CYP1A9(I391A) showed higher activities than that of CYP1A9 toward progesterone. The substrate specificity of CYP1A9 may be altered by replacing an amino acid related to substrate binding. (C) 2015 Elsevier B.V. All rights reserved.

  ELSEVIER SCIENCE BV, Sep. 2015, ENVIRONMENTAL TOXICOLOGY AND PHARMACOLOGY, 40(2) (2), 360 - 368, English

  [Refereed]

  Scientific journal


• Point mutation of cytochrome P450 2A6 (a polymorphic allele CYP2A6.25) confers new substrate specificity towards flavonoids

  UNO Tomohide, OGURA Chiaki, IZUMI Chihiro, NAKAMUJRA Masahiko, YANASE Takeshi, Hiroshi YAMAZAKI, ASHIDA Hitoshi, KANAMARU Kengo, YAMAGATA Hiroshi, IMAISHI Hiromasa

  Jul. 2015, Biopharmaceutics & Drug Disposition, 36(8) (8), 552 - 563, English

  [Refereed]

  Scientific journal


• Characterization of Rab-interacting lysosomal protein in the brain of Bombyx mori

  Tomohide Uno, Yuri Isoyama, Kazuki Sakamoto, Yuichi Uno, Katsuhiko Sakamoto, Kengo Kanamaru, Hiroshi Yamagata, Michihiro Takagi, Akira Mizoguchi, Makio Takeda

  Rab guanosine triphosphatases in eukaryotic cells are key regulators of membrane-trafficking events, such as exocytosis and endocytosis. Rab7 regulates traffic from early to late endosomes and from late endosomes to vacuoles/lysosomes. The Rab7-interacting lysosomal protein (RILP) was extracted from the silkworm, Bombyx mori (B. mori), and expressed in Escherichia coli (E. coli), followed by its purification. The glutathione sulfotransferase pull-down assay revealed that Rab7 of B. mori interacted with RILP of B. mori. We then produced antibodies against RILP of B. mori in rabbits for their use in Western immunoblotting and immunohistochemistry. Western immunoblotting of brain tissue for RILP revealed a single band, at approximately 50 kD. RILP-like immunohistochemical reactivity (RILP-ir) was restricted to neurons of the pars intercerebralis and dorsolateral protocerebrum. Furthermore, RILP-ir was colocalized with the eclosion hormone-ir and bombyxin-ir. However, RILP-ir was not colocalized with prothoracicotropic hormone-ir. These results were similar to those of Rab7 from our previous study. These findings suggest that RILP and Rab7 are involved in the neurosecretion in a restricted subtype of neurons in B. mori. Thus, our study is the first to report of a possible relationship between an insect Rab effector and neurosecretion.

  SPRINGER, Mar. 2014, HISTOCHEMISTRY AND CELL BIOLOGY, 141(3) (3), 311 - 320, English

  [Refereed]

  Scientific journal


• A cis-element responsible for cGMP in the promoter of the soybean chalcone synthase gene

  Hamad Abu Zahra, Satoru Kuwamoto, Tomohide Uno, Kengo Kanamaru, Hiroshi Yamagata

  The cyclic nucleotides cGMP and cAMP have been reported to play key roles in the regulation of plant processes and responses. We have previously reported that several genes encoding flavonoid biosynthetic enzymes, including chalcone synthase (CHS) in soybean (Glycine max L), were induced by cGMP but not cAMP. The soybean genome contains nine CHS gene copies (GmCHS1-9). We investigated the responsiveness of several GmCHS genes to cGMP, CAMP, NO, and white light. Quantitative RT-PCR analysis showed that the transcript levels of GmCHS7 and GmCHS8 were increased by 3.6- and 3.8-fold, respectively, with cGMP whereas the transcript levels of GmCHS2 remained constant. Although CAMP had no effect on the transcript levels of the three genes, NO had an activation effect on all three. White light activated the three genes in a transient manner, with GmCHS2, GmCHS7, and GmCHS8 transcript levels increasing 3-fold after 3 h and decreasing to basal levels after 9 h. The GmCHS8 promoter contains several important cis-elements, including the G-box and H-box forming the Unit-I-like sequence and the MYB binding sequence, a target of the GmMYB176 transcription factor regulating the expression of GmCHS8. A transient gene expression assay revealed the activation of the Unit-I-like sequence, but not of the MYB binding sequence, by cGMP. The combination of G-box and H-box was necessary for cGMP responsiveness. Taken together, these results suggest that the Unit-I-like sequence in the promoters of GmCHS7 and GmCHS8 is a cGMP responsive cis-element in these genes and that NO exerts its effect via cis-elements other than the Unit-I-like sequence. (C) 2013 Elsevier Masson SAS. All rights reserved.

  ELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER, Jan. 2014, PLANT PHYSIOLOGY AND BIOCHEMISTRY, 74(1) (1), 92 - 98, English

  [Refereed]

  Scientific journal


• Metabolism of 7-ethoxycoumarin, safrole, flavanone and hydroxyflavanone by cytochrome P450 2A6 variants.

  UNO TOMOHIDE, 尾部 悠一郎, Ogura Chika, 山本 貢平, KANAMARU KENGO, YAMAGATA HIROSHI, IMAISHI HIROMASA

  多種の生体異物を代謝するヒトシトクロームP450 2A6の野生型および13種の変異型酵素をNADPH-シトクロームP450還元酵素とともに大腸菌で発現し、組換え酵素の7-エトキシクマリン、サロール、フラバノン、水酸化フラバノンの代謝を解析し、2A6ファミリーがフラボノイドを代謝することを明らかにした。

  Nov. 2013, Biopharm Drug Dispos., 34(2) (2), 87 - 97, English

  [Refereed]

  Scientific journal


• Relationship between the expression of Rab family GTPases and neuropeptide hormones in the brain of Bombyx mori

  Tomohide Uno, Kazuki Sakamoto, Yuri Isoyama, Susumu Hiragaki, Yuichi Uno, Kengo Kanamaru, Hiroshi Yamagata, Michihiro Takagi, Akira Mizoguchi, Makio Takeda

  Rab proteins are small GTPases that play essential roles in vesicle transport. In this study, we examined the expression of Rab proteins and neuropeptide hormones in the brain of the silkworm, Bombyx mori. We produced antibodies against B. mori Rab1 and Rab14 in rabbits. Immunoblotting of samples of brain tissue from B. mori revealed a single band for each antibody. Rab1 and Rab14 immunohistochemical labeling in the brain of B. mori was restricted to neurons of the pars intercerebralis and dorsolateral protocerebrum. Rab1, Rab7 and Rab14 co-localized with bombyxin. Rab1 and Rab7 co-localized with eclosion hormone. Rab1 co-localized with prothoracicotropic hormone. These results suggest that Rab1, Rab7 and Rab14 may be involved in neuropeptide transport in the brain of B. mori. This is the first report on the specificity of Rab proteins for the secretion of different neuropeptides in insects.

  SPRINGER, Feb. 2013, HISTOCHEMISTRY AND CELL BIOLOGY, 139(2) (2), 299 - 308, English

  [Refereed]

  Scientific journal


• Comparison of three hemA1 cDNA sequences putatively encoding a glutamyl tRNA reductase in common wheat

  Yu Takenouchi, Kengo Kanamaru, TAKUMI SHIGEO

  Feb. 2012, Wheat Information Service, 113, 9 - 11, English

  Research society


• Characterization of Three Homoeologous cDNAs Encoding Chloroplast-targeted Aminolevulinic Acid Dehydratase in Common Wheat

  Yu Takenouchi, Haruka Nakajima, Kengo Kanamaru, Shigeo Takumi

  In the tetrapyrrole biosynthetic pathway of higher plants, 5-aminolevulinic acid (ALA) is metabolized by ALA dehydratase (ALAD). Here, we isolated ALAD1 cDNA from common wheat (Triticum aestivum L.) and its diploid progenitors, and produced transgenic tobacco plants expressing the wheat ALAD1 gene. The ALAD1 genes were highly conserved among wheat relatives, and three homoeologous loci of wheat ALAD1 (TaALAD1) were equally transcribed in common wheat. A transient expression assay of a TaALAD1-GFP (green fluorescent protein) fusion protein suggested that TaALAD1 is localized in chloroplasts. Overexpression of TaALAD1 in transgenic tobacco resulted in a significant increase in ALAD activity in leaves. Moreover, the transgenic tobacco showed vigorous growth and increased survival rate on medium containing ALA at herbicidal concentrations. These results indicate that wheat ALAD1 has catalytic activity in metabolizing ALA in plastids, and that ectopic expression of TaALAD1 in transgenic plants increases their tolerance to ALA application at high concentrations.

  WILEY-BLACKWELL, Dec. 2011, JOURNAL OF INTEGRATIVE PLANT BIOLOGY, 53(12) (12), 942 - 950, English

  [Refereed][Invited]

  Scientific journal


• Metabolism of the herbicides chlorotoluron, diuron, linuron, simazine, and atrazine by CYP1A9 and CYP1C1 from Japanese eel (Anguilla japonica)

  Tomohide Uno, Satoru Kaji, Tatsushi Goto, Hiromasa Imaishi, Masahiko Nakamura, Kengo Kanamaru, Hiroshi Yamagata, Yoshio Kaminishi, Takao Itakura

  Through the use of a number of bioconversion experiments we demonstrated that P450 proteins (CYP1A9 and CYP1C1) from Japanese eel (Anguilla japonica) metabolized a number of herbicides and the drug phenacetin. We performed bioconversion experiments in which substrates were added directly to incubation medium. The resulting metabolites were extracted and analyzed by high-performance liquid chromatography. Proteins CYP1A9 and CYP1C1 metabolized 50 nmol of the drug phenacetin to yield 12.1 and 1.1 nmol of product (acetaminophen), respectively. Further incubation of CYP1A9 with 50 nmol of the herbicides chlorotoluron, diuron, linuron, simazine, or atrazine yielded 16.5, 18.5, 7.3, 1.6, or 0.8 nmol of product, respectively. CYP1C1 also metabolized linuron, diuron, and simazine yield 5.4. 4.6, or 0.7 nmol of product, respectively. Next, polyclonal antibody was isolated by immunizing with two conjugated-peptides (amino acid residues 272-290 and 294-310) of CYP1A9. This antibody did not recognize human CYP1A2 or CYP1C1. Western blotting using the antibody revealed one band in the livers of Japanese eel and tilapia (Oreochromis niloticus). Theses results suggest that CYP1A9 and CYP1C1 metabolize herbicides, and that CYP1A9 is an useful biomarker of contamination when detected with this antibody. (C) 2011 Elsevier Inc. All rights reserved.

  ACADEMIC PRESS INC ELSEVIER SCIENCE, Oct. 2011, PESTICIDE BIOCHEMISTRY AND PHYSIOLOGY, 101(2) (2), 93 - 102, English

  [Refereed]

  Scientific journal


• Small GTPases of the Rab family in the brain of Bombyx mori

  Tomohide Uno, Keisuke Hata, Susumu Hiragaki, Yuri Isoyama, Le Thi Dieu Trang, Yuichi Uno, Kengo Kanamaru, Hiroshi Yamagata, Masahiko Nakamura, Michihiro Takagi, Makio Takeda

  Small GTPases of the Rab family are key regulators of membrane trafficking. We produced antibodies against the Rab7 protein of Bombyx mori (BRab7) in rabbits, and against the Rab11 protein of B. mori (BRab11) in mice. The antibodies recognized BRab7 and BRab11 proteins, but did not recognize other Rab proteins. Immunoblotting of samples from brain tissue of B. mori revealed a single band for each antibody. Rab11 was expressed in most tissues, whereas Rab7 was expressed in the brain, ovary, and testis. Immunohistochemical reactivity of Rab7 and Rab11 in the brain of B. mori was restricted to neurons of the pars intercerebralis and dorsolateral protocerebrum. Double-labeling experiments demonstrated that immunohistochemical reactivity of Rab7 co-localized with that of Rab11 and partially with that of Rab8. Immunohistochemical reactivity of Rab11 and Rab8 co-localized with that of PERIOD, one of the proteins associated with circadian rhythm. These findings suggest that Rab7, Rab8, and Rab11 are involved in protein transport in the neurons of the brain of B. mori and might play a role in the control of circadian rhythm.

  SPRINGER, Dec. 2010, HISTOCHEMISTRY AND CELL BIOLOGY, 134(6) (6), 615 - 622, English

  [Refereed]

  Scientific journal


• Functional Analysis of the Cucumisin Propeptide as a Potent Inhibitor of Its Mature Enzyme

  Masataka Nakagawa, Megumi Ueyama, Hiroki Tsuruta, Tomohide Uno, Kengo Kanamaru, Bunzo Mikami, Hiroshi Yamagata

  Cucumisin is a subtilisin-like serine protease (subtilase) that is found in the juice of melon fruits (Cucumis melo L.). It is synthesized as a preproprotein consisting of a signal peptide, NH(2)-terminal propeptide, and 67-kDa protease domain. We investigated the role of this propeptide (88 residues) in the cucumisin precursor. Complementary DNAs encoding the propeptides of cucumisin, two other plant subtilases (Arabidopsis ARA12 and rice RSP1), and bacterial subtilisin E were expressed in Escherichia coli independently of their mature enzymes. The cucumisin propeptide strongly inhibited cucumisin in a competitive manner with a K(i) value of 6.2 +/- 0.55 nM. Interestingly, cucumisin was also strongly inhibited by ARA12 and RSP1 propeptides but not by the subtilisin E propeptide. In contrast, the propeptides of cucumisin, ARA12, and RSP1 did not inhibit subtilisin. Deletion analysis clearly showed that two hydrophobic regions, Asn(32)-Met(38) and Gly(97)-Leu(103), in the cucumisin propeptide were important for its inhibitory activity. Site-directed mutagenesis also confirmed the role of a Val(36)-centerd hydrophobic cluster within the Asn(32)-Met(38) region in cucumisin inhibition. Circular dichroism spectroscopy revealed that the cucumisin propeptide had a secondary structure without a cognate protease domain and that the thermal unfolding of the propeptide at 90 degrees C was only partial and reversible. A tripeptide, Ile(35)-Val(36)-Tyr(37), in the Asn(32)-Met(38) region was thought to contribute toward the formation of a proper secondary structure necessary for cucumisin inhibition. This is the first report on the function and structural information of the propeptide of a plant serine protease.

  AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, Sep. 2010, JOURNAL OF BIOLOGICAL CHEMISTRY, 285(39) (39), 29797 - 29807, English

  [Refereed]

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Rab14 from Bombyx mori (Lepidoptera: Bombycidae) shows ATPase activity

  Tomohide Uno, Tsubasa Moriwaki, Yuri Isoyama, Yuichi Uno, Kengo Kanamaru, Hiroshi Yamagata, Masahiko Nakamura, Michihiro Takagi

  Rab GTPases are essential for vesicular transport, whereas adenosine triphosphate (ATP) is the most important and versatile of the activated carriers in the cell. But there are little reports to clarify the connection between ATP and Rab GTPases. A cDNA clone (Rab14) from Bombyx mori was expressed in Escherichia coli as a glutathione S-transferase fusion protein and purified. The protein bound to [H-3]-GDP and [S-35]-GTPgS. Binding of [S-35]-GTP gamma S was inhibited by guanosine diphosphate (GDP), guanosine triphosphate (GTP) and ATP. Rab14 showed GTP- and ATP-hydrolysis activity. The Km value of Rab14 for ATP was lower than that for GTP. Human Rab14 also showed an ATPase activity. Furthermore, bound [H-3]-GDP was exchanged efficiently with GTP and ATP. These results suggest that Rab14 is an ATPase as well as GTPase and gives Rab14 an exciting integrative function between cell metabolic status and membrane trafficking.

  ROYAL SOC, Jun. 2010, BIOLOGY LETTERS, 6(3) (3), 379 - 381, English

  [Refereed]

  Scientific journal


• Further Evaluation of the Localization and Functionality of Hemagglutinin Epitope- and Fluorescent Protein-Tagged AtMinD1 in Arabidopsis thaliana

  Makoto T. Fujiwara, Dongliang Li, Yusuke Kazama, Tomoko Abe, Tomohide Uno, Hiroshi Yamagata, Kengo Kanamaru, Ryuuichi D. Itoh

  Symmetric chloroplast division requires a prokaryote-derived division regulator protein MinD, whose subchloroplastic localization remains to be completely established. We investigated the localization and functionality of AtMinD1 (Arabidopsis thaliana MinD) fused with a dual hemagglutinin epitope (dHA) or a yellow fluorescent protein (YFP). AtMinD1-dHA, which successfully complemented the arc11/atminD1 mutant phenotype, was predominantly located at the envelope membrane and the mid-chloroplast constriction site. Meanwhile, AtMinD1-YFP was non-functional and showed suborganellar localization partly similar to that of AtMinD1-dHA. This prompts us to reevaluate earlier transgenic and transient expression studies using fluorescent protein-tagged AtMinD1.

  TAYLOR & FRANCIS LTD, Jul. 2009, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 73(7) (7), 1693 - 1697, English

  [Refereed]

  Scientific journal


• Biochemical characterization of Rab proteins from Bombyx mori

  Tomohide Uno, Tsubasa Moriwaki, Masahiko Nakamura, Mamoru Matsubara, Hiroshi Yamagata, Kengo Kanamaru, Michihiro Takagi

  The small GTPases known as Rab proteins are key regulators of membrane trafficking. We used RT-PCR to isolate cDNA clones of insect-specific Rab proteins (BRabN1 and BRabN2) showing low homology with known Rab proteins from other animals, from mRNA of Bombyx mori. These 2 Rabs were produced in Escherichia coli and purified. BRabN1 bound [3H]-GDP and [ 35S]-GTPγS with dissociation constants of 0.087 × 10 -6 M and 1.02 × 10-6 M, respectively, whereas those of BRabN2 were 0.546 × 10-6 M and 1.02 × 10-6 M, respectively. Binding of [35S]-GTPγS to BRabN1 and N2 was inhibited by GDP and GTP. The GTP-hydrolysis activities of BRabN1 and N2 were 154 and 35.5 mmol/min/mole, respectively, and bound [35S]-GTPγS was exchanged efficiently with GTP. BRabN1 also showed ATPase activity and exchange of [35S]-GTPγS with ATP. Monoclonal antibodies against BRabN1 and N2 did not recognize any other Rab proteins, and Western blotting using the anti-BRabN1 antibody revealed a single band in the testis of B. mori. These results suggest that BRabN1 and N2 of B. mori bind GTP, convert from the GTP-bound state to the GDP-bound state by intrinsic GTP hydrolysis activity, and return to the GTP-bound state with the exchange, and that BRabN1 is specifically expressed in testis. © 2008 Wiley Periodical Inc.

  Feb. 2009, Archives of Insect Biochemistry and Physiology, 70(2) (2), 77 - 89, English

  [Refereed]

  Scientific journal


• Phosphorylation of small GTPase rab proteins from Bombyx mori (Lepidoptera: Bombycidae)

  Tomohide Uno, Keisuke Hata, Le Thi Dieu Trang, Susumu Hiragaki, Takuya Nakada, Masahiko Nakamura, Yuichi Uno, Hiroshi Yamagata, Kengo Kanamaru, Makio Takeda, Mamoru Matsubara

  Small GTPases of the Rab family act as essential regulators of vesicle transport pathways. Five Rab cDNA clones (BRab1, 7, 8, 11 and 14) from Bombyx mori were expressed in Escherichia coli as a thioredxin or glutathione sulfotransferase fusion protein. After purification, the fusion protein was tested for phosphorylation using protein kinase C (PKC). Results indicate that all of them were phosphorylated in vitro. The phosphorylation site of BRab1 was determined by mass-spectrometric analysis, which identified that Ser-17 of BRab1 was phosphorylated by PKC. Deletion and site-directed mutagenesis indicated that Ser-111of BRab8, in addition to Ser-17, was newly phosphorylated. Further immunohistochemical analysis using antibodies against Rab8 indicated that there are some Rab8 immunoreactive cells close to the neuropeptide secreting cells. This result suggests that in insects Rab proteins are regulated by phosphorylation and at least some of them are involved in neuropeptide secretion.

  Czech Academy of Sciences, 2009, European Journal of Entomology, 106(4) (4), 499 - 506, English

  [Refereed]

  Scientific journal


• Ferulic Acid Production in the Brewing of Rice Wine (Sake)

  Tomohide Uno, Atsushi Itoh, Tetsuya Miyamoto, Masaharu Kubo, Kengo Kanamaru, Hiroshi Yamagata, Yukio Yasufuku, Hiromasa Imaishi

  The traditional Japanese alcoholic beverage sake is produced by fermentation of rice by Saccharomyces cerevisiae and Aspergillus oryzae. A. oryzae releases ferulic acid. an antioxidant, from steamed rice during the fermentation process. The concentration of ferulic acid increased with time during fermentation and the production rate peaked 9-12 days post inoculation. Analysis of the fermentation cultures of Aspergillus oryzae, by high-performance liquid chromatography (HPLC), revealed that p-coumaric acid induced an 18.9-fold increase in the level of ferulic acid. Furthermore, SDS-PAGE analysis revealed an increase or decrease in the level of specific proteins after the addition of p-coumaric acid to fermentation Cultures of Aspergillus oryzae. Ferulate esterase (FAE) activity was observed in the fermented sake ten days following the start of the fermentation process. These results suggest that the level of ferulic acid is regulated by the enzymes synthesized by A. oryzae during the sake brewing process.

  INST BREWING, 2009, JOURNAL OF THE INSTITUTE OF BREWING, 115(2) (2), 116 - 121, English

  [Refereed]

  Scientific journal


• Cyclic GMP acts as a common regulator for the transcriptional activation of the flavonoid biosynthetic pathway in soybean

  Kenji Suita, Takaaki Kiryu, Maki Sawada, Maiko Mitsui, Masataka Nakagawa, Kengo Kanamaru, Hiroshi Yamagata

  Cyclic GMP (cGMP) is an important signaling molecule that controls a range of cellular functions. So far, however, only a few genes have been found to be regulated by cGMP in higher plants. We investigated the cGMP-responsiveness of several genes encoding flavonoid-biosynthetic enzymes in soybean (Glycine max L.) involved in legume-specific isoflavone, phytoalexin and anthocyanin biosynthesis, such as phenylalanine ammonia-lyase, cinnamate 4-hydroxylase, 4-coumarate:CoA ligase, chalcone synthase, chalcone reductase, chalcone isomerase, 2-hydroxyisoflavanone synthase, 2-hydroxyisoflavanone dehydratase, anthocyanidin synthase, UDP-glucose:isoflavone 7-O-glucosyltransferase, and isoflavone reductase, and found that the majority of these genes were induced by cGMP but not by cAMP. All cGMP-induced genes were also stimulated by sodium nitroprusside (SNP), a nitric oxide (NO) donor, and illumination of cultured cells with white light. The NO-dependent induction of these genes was blocked by 6-anilino-5,8-quinolinedione, an inhibitor of guanylyl cyclase. Moreover, cGMP levels in cultured cells were transiently increased by SNP. Consistent with the increases of these transcripts, the accumulation of anthocyanin in response to cGMP, NO, and white light was observed. The treatment of soybean cotyledons with SNP resulted in a high accumulation of isoflavones such as daidzein and genistein. Loss- and gain-of-function experiments with the promoter of chalcone reductase gene indicated the Unit I-independent activation of gene expression by cGMP. Together, these results suggest that cGMP acts as a second messenger to activate the expression of genes for enzymes involved in the flavonoid biosynthetic pathway in soybean.

  SPRINGER, Jan. 2009, PLANTA, 229(2) (2), 403 - 413, English

  [Refereed]

  Scientific journal

  Link to Kobe Univ. Repository (Kernel)


• Bioconversion of small molecules by cytochrome P450 species expressed in Escherichia coli

  Tomohide Uno, Sota Okamoto, Satoko Masuda, Atsushi Itoh, Yuichi Uno, Masahiko Nakamura, Kengo Kanamaru, Hiroshi Yamagata, Hiromasa Imaishi

  P450 (cytochrome P450) enzymes catalyse the mono-oxygenation of a wide range of compounds such as steroids, fatty acids, vitamins and drugs. In the present paper we demonstrate a system for bioconverting diverse compounds [flavanone, DHEA (dehydroepiandrosterone) and 7-ethoxycoumarin] using P450 species expressed in Escherichia coli. First, we expressed four P450 species: rabbit CYP2B (MO family 2, subfamily B), fruitfly (Drosophila) CYP317A, rat CYP3A23 and mouse CYP2J5. Next, we added substrates directly to the incubation medium. The resulting metabolites were extracted and analysed by HPLC and spectrofluorimetry. The first substrate, 7-ethoxycoumarin, was de-ethylated by CYP2B; CYP2J5 and CYP3A23 showed weak activity, and CYP317A had no activity for 7-ethoxycoumarin. We next used flavanone, a flavonoid, as a substrate for these four MO species and other P450 species expressed previously. As a result, CYP2B, CYP2C43 and CYP2C29 catalysed flavanone 2-hydroxylation. CYP2A5 catalysed 2- and 4-hydroxylations. Finally, to produce diverse modified compounds, variants of CYP2A5 with point mutations were incubated with a steroid (DHEA) and an antioxidant (flavanone) in vivo. HPLC analysis indicated that two P450 species produced a 7-beta-hydroxy-DHEA and two P450 species produced a 2-alpha-hydroxy-DHEA. Four P450 species catalysed flavanone 2- and 4-hydroxylations. These results indicate that bioconversion by P450 is a useful technique to modify small molecules (steroids, coumarin and flavanone) and produce new, diverse hydroxylated compounds, which could be used for high-throughput screening for drug discovery.

  WILEY, Aug. 2008, BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY, 50, 165 - 171, English

  [Refereed]

  Scientific journal


• Bioconversion by functional P450 1A9 and P450 1C1 of Anguilla japonica

  Tomohide Uno, Sota Okamoto, Satoko Masuda, Hiromasa Imaishi, Masahiko Nakamura, Kengo Kanamaru, Hiroshi Yamagata, Mohamed A. H. El-Kady, Yoshio Kaminishi, Takao Itakura

  We indicated that two P450s (1A9 and 1C1) from Japanese eel (Anguilla japonica) metabolized 7-ethoxycoumarin, 7-ethoxyresorufin, and flavanone. At first, we constructed expression vectors for two types of P450 (1A9 and 1C1). The reduced CO-difference spectra of Escherichia coli cells transformed with these plasmids showed Soret peaks (450 nm) that were typical of P450s. We performed bioconversion experiments in which substrates were added directly to incubation medium. The resulting metabolite(s) were extracted and analyzed by high-performance liquid chromatography and spectrofluorometer. Incubation of 50 nmol 7-ethoxyresorufin with P450 1C1 yielded 0.773 nmol of deethylated product, whereas 50 nmol 7-ethoxycoumarin resulted in 4.76 nmol. P450 1A9 metabolized 50 nmol of 7-ethoxyresorufin and 7-ethoxycoumarin to yield 6.54 and 20.9 nmol of deethylated product, respectively. Incubation of 50 nmol flavanone with P450 1C1 yielded 1.46 nmol and 0.69 nmol of products, whereas 50 mnol flavanone with P450 1A9 resulted in 1.10 nmol. In this system, 4'-hydroxy flavanones were formed by P450 1A9 and P450 1C1. P450 1A9 also metabolized 50 nmol of 17 beta-estradiol to yield 4.25 nmol of product. In this system, 2-hydroxy estradiol was formed by P450 1A9 using 17 beta-estradiol as a substrate. This study is the first to identify the substrates that P450 1C1 and 1A9 metabolize. (c) 2007 Elsevier Inc. All rights reserved.

  ELSEVIER SCIENCE INC, Apr. 2008, COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY C-TOXICOLOGY & PHARMACOLOGY, 147(3) (3), 278 - 285, English

  [Refereed]

  Scientific journal


• Monoclonal antibody against rab8 from Bombyx mori (Lepidoptera : Bombycidae)

  Tomohide Uno, Takuya Nakada, Yuichi Uno, Kengo Kanamaru, Hiroshi Yamagata, Masahiko Nakamura, Michihiro Takagi

  Small GTPases of the Rab family are key regulators of membrane trafficking. Monoclonal antibodies are useful tools for identifying proteins that interact with other proteins and for examining their tissue distribution. We selected a monoclonal antibody against Rab8 of Bombyx Mori L. It specifically recognized amino acid residues 30-109, which are conserved among Rab8 proteins, and did not recognize any other Rab proteins. Western blotting using the antibody revealed one band in the brains of B. Mori and rat. Far-Western blotting analysis detected three proteins interacting with Rab8. These results indicate that this antibody is useful for clarifying the physiological function of Rab8 of B. Mori and other species. This is a report of a study on a monoclonal antibody against insect Rab protein.

  CZECH ACAD SCI, INST ENTOMOLOGY, Oct. 2007, EUROPEAN JOURNAL OF ENTOMOLOGY, 104(4) (4), 641 - 645, English

  [Refereed]

  Scientific journal


• Determination of phosphorylated amino acid residues of Rab8 from Bombyx mori

  Tomohide Uno, Takuya Nakada, Sota Okamaoto, Masahiko Nakamura, Mamoru Matsubara, Hiromasa Imaishi, Hiroshi Yamagata, Kengo Kanamaru, Michihiro Takagi

  The Rob family of small GTPases are key regulators of membrane trafficking. Partially purified ROB from Bombyx mori (BRab8) was phosphorylated by protein kinase C in mammalian cells in vitro. To determine which of the seven serines and four threonines are phosphorylated, we generated deletion and site-directed mutants of BRab8, inserted them in Escherichia coli, partially purified the encoded fusion proteins by affinity chromatography, and examined their phosphorylation by protein kinase C in vitro. We found that Ser-132 of BRab8 was specifically phosphorylated by protein kinase C. In addition, Western blotting using an antiserum against BRab8 and in-gel staining for phosphorylated proteins revealed that BOB is phosphorylated in vivo.

  WILEY-BLACKWELL, Oct. 2007, ARCHIVES OF INSECT BIOCHEMISTRY AND PHYSIOLOGY, 66(2) (2), 89 - 97, English

  [Refereed]

  Scientific journal


• Coordinated functions of plastid sigma factors in chloroplast development and gene expression

  T. Shiina, Y. Ishizaki, K. Ozono, C. Takenaka, M. Hanaoka, K. Kanamaru, K. Tanaka, Y. Nakahira

  SPRINGER, Feb. 2007, PHOTOSYNTHESIS RESEARCH, 91(2-3) (2-3), 267 - 267, English


• Cloning and characterization of soybean GmGT-1 that binds to a light-responsive cis-element in ELIP promoter.

  Aoki, M, Uno, T, Kanamaru, K, Yamagata, H

  2007, Kobe Univ. Frontier Technology Forum 2007, 17, English

  [Refereed]

  Symposium


• Development and characterization of photo-autophilic Arabidopsis cultured cells

  Li, T, Uno, T, Yamagata, H, Kanamaru, K

  2007, Kobe Univ. Frontier Technology Forum 2007, 18, English

  [Refereed]

  Symposium


• Small GTP binding proteins; Rab GTPases from Bombyx mori.

  Moriwaki, T, Nakada, T, Uno, T, Kanamaru, K, Yamagata, H

  2007, Kobe Univ. Frontier Technology Forum 2007, 19

  [Refereed]


• Bioconversion by functional P450 1A9 and 1C1of Anguillus japonica,

  UNO TOMOHIDE, Sota Okamoto, Satoko Masuda, Hiromasa Imaishi, Masahiko Nakamura, Kengo Kanamaru, Hiroshi Yamagata, Ei-Kady, Kaminishi, Takao Itakura

  2007, Comprehensive biochemistry and physiology C Toxicology Pharamacology, 147, 278 - 285, English

  [Refereed]

  Scientific journal


• Modification of small molecules by using cytochrome P450 expressed in Escherichia coli

  Tomohide Uno, Atsushi Nakao, Satoko Masuda, Yuuki Taniguchi, Kengo Kanamaru, Hiroshi Yamagata, Masahiko Nakamura, Hiromasa Imaishi, Kiyoharu Oono

  We developed a system for bioconverting diverse compounds using P450s produced in Escherichia coli. Vectors for the expressing various P450 cDNAs quickly and easily in E. coli were developed by using several restriction enzyme sites. Three types of P450 (2C2, 2C29, and 2D22) were produced using these plasmids. Substrates were directly added to the incubation medium and metabolized. To obtain pure product from the medium, we first tried production of P450 in synthetic medium. The amount of another P450 2C43 produced in the synthetic medium was similar to the amount produced in Luria broth (LB) medium. Next, estradiol, a steroid, was added as a substrate, incubated, and the metabolite was extracted and analyzed by high-performance liquid chromatography. The metabolite extracted from synthetic medium was purer than that obtained from LB medium. Three P450s (2C29, 2C2, and 2A4) metabolized testosterone at different positions. P450 2C29 metabolized 7-ethoxycournarin, androstendione, and dehydroepiandrosterone in this medium. P450s produced in the synthetic medium may be useful for producing various modified compounds for high-throughput screening.

  SPRINGER HEIDELBERG, Dec. 2006, JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY, 33(12) (12), 1043 - 1050, English

  [Refereed]

  Scientific journal


• Rice bifunctional alpha-amylase/subtilisin inhibitor: Cloning and characterization of the recombinant inhibitor expressed in Escherichia coli

  Teruyuki Yamasaki, Masaki Deguch, Toshiko Fujimoto, Takehiro Masumura, Tomohide Uno, Kengo Kanamaru, Hiroshi Yamagata

  The complete nucleotide sequences of the cDNA and its gene that encode a bifunctional alpha-amylase/subtilisin inhibitor of rice (Oryza sativa L.) (RASI) were analyzed. RASI cDNA (939 lip) encoded a 200-residue polypeptide with a molecular mass of 21,417 Da, including a signal peptide of 22 amino acids. Sequence comparison and phylogenetic analysis showed that RASI is closely related to alpha-amylase/subtilisin inhibitors from barley and wheat. RASI was found to be expressed only in seeds, suggesting that it has a seed-specific function. A coding region of RASI cDNA without the signal peptide was introduced into Escherichia coli and was expressed as a His-tagged protein. Recombinant RASI was purified to homogeneity in a single step by Ni-chelating affinity column chromatography and characterized to elucidate the target enzyme. The recombinant inhibitor had strong inhibitory activity toward subtilisin, with an equimolar relationship, comparable with that of native RASI, and weak inhibitory activity toward some microbial alpha-amylases, but not toward animal or insect alpha-amylases. These results suggest that RASI might function in the defense of the seed against microorganisms.

  TAYLOR & FRANCIS LTD, May 2006, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 70(5) (5), 1200 - 1209, English

  [Refereed]

  Scientific journal


• Biochemical characterization of Rab proteins from Bombyx mori,

  UNO TOMOHIDE, Tsubasa Moriwaki, Masahiko Nakamura, Mamoru Matsubara, Hiroshi Yamagata, Kengo Kanamaru, Michihiro Takagi

  The small GTPases known as Rab proteins are key regulators of membrane trafficking. We used RT-PCR to isolate cDNA clones of insect-specific Rab proteins (BRabN1 and BRabN2) showing low homology with known Rab proteins from other animals, from mRNA of Bombyx mori. These 2 Rabs were produced in Escherichia coli and purified. BRabN1 bound [(3)H]-GDP and [(35)S]-GTPgammaS with dis

  2006, Archives of Insect Biochemistry and Physiology, 70, 77 - 89, English

  [Refereed]

  Scientific journal


• Expression, purification and characterization of methyl DNA binding protein from Bombyx mori.

  Uno T, Nomura Y, Nakamura M, Nakao A, Tajima S, Kanamaru K, Yamagata H, Iwanaga Y

  Aug. 2005, J Insect Sci, 5:8, 1-8

  [Refereed]

  Scientific journal


• Glutamyl-tRNA mediates a switch in RNA polymerase use during chloroplast biogenesis.

  Hanaoka M, KANAMARU KENGO, Fujiwara M, Takahashi H, Tanaka K

  Chloroplast genes of higher plants are transcribed by two types of RNA polymerase that are encoded by nuclear (NEP (nuclear-encoded plastid RNA polymerase)) or plastid (PEP (plastid-encoded plastid RNA polymerase)) genomes. NEP is largely responsible for the transcription of housekeeping genes during early chloroplast development. Subsequent light-dependent chloroplast maturati

  Jun. 2005, EMBO Rep., 6(6) (6), 545 - 50, English, International magazine

  [Refereed]

  Scientific journal


• Roles of chloroplast RNA polymerase sigma factors in chloroplast development and stress response in higher plants

  K Kanamaru, K Tanaka

  Chloroplast transcription in higher plants is performed by two types of RNA polymerases, plastidencoded RNA polymerase (PEP) and nuclear-encoded RNA polymerase (NEP). PEP is a eubacteria-type multisubunit enzyme whose catalytic core subunits are encoded by the chloroplast genome, whereas NEP is the nuclear encoded T7 phage-type single subunit enzyme. PEP is critical for the biogenesis and maintenance of chloroplasts, and is finely tuned by the nuclear encoded sigma subunits. Of the six Arabidopsis sigma subunits, SIG2 is involved in the transcription of several chloroplast tRNA genes, including trnE encoding tRNA-Glu. SIG2 possibly couples translation and pigment synthesis in chloroplasts. On the other hand, SIG5 is induced by various stresses and contributes to repair of damaged photosystem 11 (PS11) through transcription of the psbD and psbC genes. Thus target genes and the physiological role of each sigma subunit are becoming clearer.

  TAYLOR & FRANCIS LTD, Nov. 2004, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 68(11) (11), 2215 - 2223, English

  [Refereed]


• DNA microarray analysis of plastid gene expression in an Arabidopsis mutant deficient in a plastid transcription factor sigma, SIG2

  A Nagashima, M Hanaoka, R Motohashi, M Seki, K Shinozaki, K Kanamaru, H Takahashi, K Tanaka

  The plastid genome of higher plants contains more than one hundred genes for photosynthesis, gene expression, and other processes. Plastid transcription is done by two types of RNA polymerase, PEP and NEP. PEP is a eubacteria-type RNA polymerase that is essential for chloroplast development. In Arabidopsis thaliana, six sigma factors (SIG1-6) are encoded by the nuclear genome, and postulated to determine the transcription specificity of PEP. In this study, we constructed a DNA microarray for all of the plastid protein-coding genes, and analyzed the effects of the sig2 lesion on the global plastid gene expression. Of the 79 plastid protein genes, it was found that only the psaJ transcript was decreased in the mutant, whereas transcripts of 47 genes were rather increased. Since many of the upregulated genes are under the control of NEP, it was suggested that the NEP activity was increased in the sig2-1 mutant.

  TAYLOR & FRANCIS LTD, Mar. 2004, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 68(3) (3), 694 - 704, English

  [Refereed]

  Scientific journal


• Analysis of the chloroplast RNA polymerase sigma factor, SIG5, in Arabidopsis thaliana: Roles in the stress-responsive chloroplast transcription.

  Nagashima A, Hanaoka M, Fujiwara M, Shikanai T, Kanamaru K, Takahashi H, Tanaka K

  The plastid genome contains more than one hundred genes and the expression is regulated depending on plastid types and environmental conditions. Sigma factors determine the specificity of the eubacteria-type plastid RNA polymerase (PEP), and the roles of sigma factors for chloroplast gene expression remain largely unknown. In this study, we analyzed the expression of the sigma factors (SIG1-6) under various stress conditions, and found that the SIG5 transcript was rapidly and drastically induced under various stresses conditions. Because this induction was well correlated with the activation of the light-responsive promoter (LRP) of psbD, and because the psbD-LRP activation was abolished in the newly identified sig5 mutant (sig5-2), we conclud that SIG5 is a stress-responsive sigma factor in Arabidopsis, and responsible for the psbD-LRP transcription. We also found that the restoration of PSII activity after high light irradiation was delayed in the sig5 mutant, suggesting roles of SIG5 under stress conditions.

  The Japanese Society of Plant Physiologists, 2004, Plant and Cell Physiology, 45, S33 - 23

  [Refereed]


• Function of a neclear-encoded sigma factor in chloroplasts; SIG2-dependent expression of some plastid-encoded tRNA genes including trnE in Arabidopsis thaliana.

  KANAMARU Kengo, TANAKA K

  2004, Endocytobiosis and Cell Research, 15,218-234, English

  [Refereed]

  Scientific journal


• The multiple-stress responsive plastid sigma factor, SIG5, directs activation of the psbD blue light-responsive promoter (BLRP) in Arabidopsis thaliana

  A Nagashima, M Hanaoka, T Shikanai, M Fujiwara, K Kanamaru, H Takahashi, K Tanaka

  Transcription in higher plant plastids is performed by two types of RNA polymerases called NEP and PEP, and expression of photosynthesis genes in chloroplasts is largely dependent on PEP, a eubacteria-type multi-subunit enzyme. The transcription specificity of PEP is modulated by six nuclear-encoded sigma factors (SIG1 to SIG6) in Arabidopsis thaliana. Here, we show that one of the six sigma factors, SIG5, is induced under various stress conditions, such as high light, low temperature, high salt and high osmotic conditions. Interestingly, transcription from the psbD blue light-responsive promoter (psbD-BLRP) was activated by not only light but also various stresses, and the transcription and the transcriptional activation of psbD-BLRP were abolished in a sig5-2 mutant. This suggests that the PEP holoenzyme containing SIG5 transcribes the psbD-BLRP in response to multiple stresses. Since the seed germination under saline conditions and recovery from damage to the PSII induced by high light were delayed in the sig5-2 mutant, we postulate that SIG5 protects plants from stresses by enhancing repair of the PSII reaction center.

  OXFORD UNIV PRESS, 2004, PLANT AND CELL PHYSIOLOGY, 45(4) (4), 357 - 368, English

  [Refereed]

  Scientific journal


• Molecular genetic analysis of chloroplast gene promoters dependent on SIG2, a nucleus-encoded sigma factor for the plastid-encoded RNA polymerase, in Arabidopsis thaliana.

  Hanaoka M, KANAMARU KENGO, Takahashi H, Tanaka K

  Most photosynthesis-related genes in mature chloroplasts are transcribed by a eubacterial-type RNA polymerase (PEP) whose core subunits are encoded by the plastid genome. It has been shown previously that six putative nuclear genes (SIG1 to SIG6) encode promoter-specificity factors for PEP in Arabidopsis thaliana, and we isolated a T-DNA insertion line of SIG2 (sig2-1 mutant) t

  Dec. 2003, Nucleic Acids Res., 31(24) (24), 7090 - 8, English, International magazine

  [Refereed]

  Scientific journal


• An Arabidopsis sigma factor (SIG2)-dependent expression of plastid-encoded tRNAs in chloroplasts

  K Kanamaru, A Nagashima, M Fujiwara, H Shimada, Y Shirano, K Nakabayashi, D Shibata, K Tanaka, H Takahashi

  A eubacteria-type RNA polymerase (PEP) plays crucial roles for chloroplast development in higher plants. The core subunits are encoded on plastid DNA (rpo genes) while the regulatory sigma factors are encoded on the nuclear DNA (SIG genes). However, the definite gene specificity of each sigma factor is unknown. We recently identified an Arabidopsis recessive pale-green mutant abc1 in which TDNA is inserted in SIG2 (sigB). In this mutant, almost normal etioplasts were developed under dark conditions while the small chloroplasts with poor thylakoid membranes and stacked lamellar were developed under light conditions. The sig2-1 mutant was deficient in accumulating enough photosynthetic and photosynthesis-related proteins as well as chlorophyll. However, mRNAs of their structural genes were not significantly reduced. Further analyses revealed that several plastid-encoded tRNAs including trnE-UUC that has dual function for protein and ALA biosyntheses were drastically reduced in the sig2-1 mutant. In contrast, nucleus-encoded T7 phage-type RNA polymerase (TNEP)dependent gene transcripts were steadily accumulated in the mutant. These results indicate that progress of chloroplast development requires SIG2-dependent expression of plastid genes, particularly some of the tRNA genes.

  OXFORD UNIV PRESS, Oct. 2001, PLANT AND CELL PHYSIOLOGY, 42(10) (10), 1034 - 1043, English

  [Refereed]

  Scientific journal


• SdiA, an Escherichia coli homologue of quorum-sensing regulators, controls the expression of virulence factors in enterohaemorrhagic Escherichia coli O157 : H7

  K Kanamaru, K Kanamaru, Tatsuno, I, T Tobe, C Sasakawa

  The quorum-sensing system in bacteria is a well-known regulatory system that controls gene expression in a cell density-dependent manner, A transcriptional regulator (LuxR homologue), signal synthase (LuxI homologue) and autoinducer (acyl homoserine lactone) are indispensable for this system in most Gram-negative bacteria. In this study, we found that SdiA, an Escherichia coli LuxR homologue, is a negative regulator of the expression of virulence factors EspD and intimin in enterohaemorrhagic E. coli (EHEC) O157:H7, The expression of EspD and intimin was inhibited at the RNA level upon SdiA overexpression, SdiA has a DNA-binding motif in its C-terminal part and can bind to the promoter regions of the esp and eae genes in vitro. Extracellular factors, which accumulate in culture supernatants of O157:H7 at the stationary phase of growth and inhibit EspD and intimin synthesis, bind to the N-terminal part of SdiA in vivo and in vitro, O157:H7 overproducing the N-terminal part of SdiA exhibited hypertranscription of EspD and intimin, suggesting that the overproduced N-terminal part had inhibited the activity of intact SdiA through titration of the extracellular factors, These results indicate that a quorum-sensing system including the SdiA protein controls colonization by O157:H7.

  WILEY-BLACKWELL, Nov. 2000, MOLECULAR MICROBIOLOGY, 38(4) (4), 805 - 816, English

  [Refereed]

  Scientific journal


• Regulation of virulence factors of enterohemorrhagic Escherichia coli O157 : H7 by self-produced extracellular factors

  K Kanamaru, K Kanamaru, Tatsuno, I, T Tobe, C Sasakawa

  Enterohemorrhagic Escherichia call (EHEC) O157:H7 causes serious diarrhea and hemolytic uremic syndrome in humans, The expressions of EspD and intimin by O157:H7 have now been shown to be downregulated by medium conditioned by O157:H7 grown at stationary phase. Preparation of conditioned medium showing the effect on the amount of EspD was not dependent on temperature or growth medium, but was dependent on growth phase. Inhibition of EspD and intimin expression was also induced by medium conditioned by E. coli K-12 strains and homoserine lactone, a signal molecule of the quorum-sensing system in Gram-negative bacteria, These results suggest the possibility that the quorum-sensing system mediated by self-produced extracellular factors plays an important role in control of colonization of EHEC O157:H7.

  TAYLOR & FRANCIS LTD, Nov. 2000, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 64(11) (11), 2508 - 2511, English

  [Refereed]

  Scientific journal


• Chloroplast development in Arabidopsis thaliana requires the nuclear-encoded transcription factor Sigma B

  Y Shirano, H Shimada, K Kanamaru, M Fujiwara, K Tanaka, H Takahashi, K Unno, S Sato, S Tabata, H Hayashi, C Miyake, A Yokota, D Shibata

  Development of plastids into chloroplasts, the organelles of photosynthesis, is triggered by light. However, little is known of the factors involved in the complex coordination of light-induced plastid gene expression, which must be directed by both nuclear and plastid genomes, We have isolated an Arabidopsis mutant, abc1, with impaired chloroplast development, which results in a pale green leaf phenotype, The mutated nuclear gene encodes a sigma factor, SigB, presumably for the eubacterial-like plastid RNA polymerase, Our results provide direct evidence that a nuclear-derived prokaryotic-like SigB protein, plays a critical role in the coordination of the two genomes for chloroplast development. (C) 2000 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.

  ELSEVIER SCIENCE BV, Nov. 2000, FEBS LETTERS, 485(2-3) (2-3), 178 - 182, English

  [Refereed]

  Scientific journal


• Chloroplast targeting, distribution and transcriptional fluctuation of AtMinD1, a eubacteria-type factor critical for chloroplast division

  K Kanamaru, M Fujiwara, M Kim, A Nagashima, E Nakazato, K Tanaka, H Takahashi

  In Arabidopsis thaliana, a mature mesophyll cell contains approximately 100 chloroplasts. Although 12 are mutants (accumulation and replication of chloroplasts) and two chloroplast division genes homologous to eubacterial ftsZ have been isolated from A, thaliana, the molecular mechanism underlying the chloroplast division is still unclear, We characterized AtMinD1, a eubacterial minD homolog, for chloroplast division in A. thaliana, AtMinD1-green fluorescent protein targeted to the chloroplasts and possibly associated with the envelope membranes in vivo. During the seed germination, the AtMinD1 transcripts were accumulated twice, just after release from cold treatment and at the beginning of rapid greening, in similar fashion to AtFtsZs. Furthermore the transcript level in a severest chloroplast division mutant, arc6, was 3-5-fold higher than that in wild-type.

  JAPANESE SOC PLANT PHYSIOLOGISTS, Oct. 2000, PLANT AND CELL PHYSIOLOGY, 41(10) (10), 1119 - 1128, English

  [Refereed]

  Scientific journal


• Three new nuclear genes, sigD, sigE and sigF, encoding putative plastid RNA polymerase sigma factors in Arabidopsis thaliana

  M Fujiwara, A Nagashima, K Kanamaru, K Tanaka, H Takahashi

  Three new nuclear genes (sigD, SigE and sigF) of Arabidopsis thaliana, encoding putative plastid RNA polymerase a factors, were identified and analyzed. Phylogenetic analysis revealed that higher plant a factors fell into at least four distinct subgroups within a diverse protein family. In addition, Arabidopsis sig genes contained conserved chromosomal intron sites, indicating that these genes arose ba DNA duplication events during plant evolution. Transcript analyses revealed two alternatively spliced transcripts generated from the sigD region, one of which is predicted to encode a a protein lacking the carboxyterminal regions 3 and 4. Finally, the amino-terminal sequence of the sigF gene product was shown to function as a plastid-targeting signal using green fluorescent protein fusions, (C) 2000 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.

  ELSEVIER SCIENCE BV, Sep. 2000, FEBS LETTERS, 481(1) (1), 47 - 52, English

  [Refereed]

  Scientific journal


• Plastidic RNA polymerase sigma factors in Arabidopsis

  K Kanamaru, M Fujiwara, M Seki, T Katagiri, M Nakamura, N Mochizuki, A Nagatani, K Shinozaki, K Tanaka, H Takahashi

  In plant cells, plastid DNA is transcribed by at least two types of RNA polymerase, plastid-encoded RNA polymerase (PEP) and nuclear-encoded RNA polymerase (NEP), PEP is homologous to eubacterial transcription machinery, but its regulatory subunit, sigma (sigma) factor, is not encoded on the plastid DNA, We previously cloned the three nuclear-encoded sigma factor genes from Arabidopsis thaliana and designated them as sigA, sigB, and sigC. By means of RFLP mapping, sigA and sigB,were mapped on chromosome I and sigC on the chromosome III. Based on comparison of the genomic structure of the three sig genes, intron sites in the 3' half of the genes were shown to be identical between sigB and sigC but divergent in sigA, consistent with the phylogenetic relevance of the three gene products. A transient expression assay of GFP fusions in Arabidopsis protoplasts showed that the N-termini of all three sig gene products functioned as chloroplast-targeting signals. We also constructed transgenic Arabidopsis lines harboring the sigA-promoter or the sigB-promoter uidA fusion. Both the sigA- and sigB-promoters were similarly activated at cotyledons, hypocotyls, rosette leaves, cauline leaves, sepals, and siliques but not at roots, seeds, or other Bower organs. In addition, the two promoters were repeatedly activated in young seedlings under continuous light, possibly in an oscillated fashion.

  JAPANESE SOC PLANT PHYSIOLOGISTS, Aug. 1999, PLANT AND CELL PHYSIOLOGY, 40(8) (8), 832 - 842, English

  [Refereed]

  Scientific journal


• Ser-534 in the hinge 1 region of Arabidopsis nitrate reductase is conditionally required for binding of 14-3-3 proteins and in vitro inhibition

  K Kanamaru, RC Wang, WP Su, NM Crawford

  14-3-3 proteins bind to the hinge 1 region of nitrate reductase (NR) and inhibit its activity. To determine which residues of NR are required for 14-3-3-inhibitory interactions, wild-type and mutant forms of Arabidopsis NR were examined in the yeast two-hybrid system and in vitro inhibition assays, NR fragments with or without hinge 1 were introduced into yeast with one of seven Arabidopsis 14-3-3 isoforms (called GF14s), NR fragments (residues 1-562 or 487-562) containing hinge 1 interacted with all GF-14s tested; an NR fragment (residues 1-487) lacking hinge 1 did not. GF14 binding to NR fragments was dependent on Ser-534, since Asp or Ala substitutions at this site blocked the interaction. Revertants with second site substitutions restoring interaction between GF14 omega and the Ala- or Asp-substituted NR fragments were identified. One isolate had a Lys to Glu substitution at position 531, which is in hinge 1, and six isolates had lie to Leu or Phe substitutions at 561 in the heme binding region. Double mutant forms of holo-NR (S534D plus K531E, I561F, or I561L) were constructed and found to be partially inhibited by protein extracts from Arabidopsis containing 14-3-3 proteins. Wild-type NR is phosphorylated and inhibited by these extracts, but S534D single mutant forms are not. These results show that inhibitory NR/14-3-3 interactions are dependent on Ser-534 but only in the context of the wild-type sequence, since substitutions at second sites render 14-3-3 binding and in vitro NR inhibition independent of Ser-534.

  AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, Feb. 1999, JOURNAL OF BIOLOGICAL CHEMISTRY, 274(7) (7), 4160 - 4165, English

  [Refereed]

  Scientific journal


• Analysis of wild-type and mutant plant nitrate reductase expressed in the methylotrophic yeast Pichia pastoris

  WP Su, JA Mertens, K Kanamaru, WH Campbell, NM Crawford

  Recombinant Arabidopsis thaliana NADH:nitrate reductase (NR; EC 1.6.6.1) was produced in the methylotrophic yeast Pichia pastoris and purified to near-electrophoretic homogeneity. purified enzyme had the spectral and kinetic properties typical of highly purified NR from natural plant sources. Site-directed mutagenesis altering several key residues and regions was carried out, and the mutant enzyme forms were expressed in P. pastoris. When the invariant cysteine residue, cysteine-191, in the molybdo-pterin region of the A. thaliana NIA2 protein was replaced with serine or alanine, the NR protein was still produced but was inactive, showing that this residue is essential for enzyme activity. Deletions or substitutions of the conserved N terminus of NR retained activity and the ability to be inactivated in vitro when incubated with ATE. Enzyme with a histidine sequence appended to the N terminus was still active and was easily purified using metal-chelate affinity chromatography. These results demonstrate that P. pastoris is a useful and reliable system for producing recombinant holo-NR from plants.

  AMER SOC PLANT BIOLOGISTS, Nov. 1997, PLANT PHYSIOLOGY, 115(3) (3), 1135 - 1143, English

  [Refereed]

  Scientific journal


• A novel member of the cspA family of genes that is induced by cold shock in Escherichia coli

  K Nakashima, K Kanamaru, T Mizuno, K Horikoshi

  Escherichia coli contains a major cold shock protein, CspA (or CS7.4), whose production is predominantly induced at low temperatures, This bacterium is known to possess five additional genes, each encoding a protein highly similar to CspA (referred to as the CspA family), Here we identified a gene that encodes a cold-shock-inducible analog of CspA and CspB, This newly cloned cspG gene is located at 22 min on the E. coli genetic map, apart from the other cspA family genes, Its gene product (70 amino acids) is 73 and 77% identical to CspA (70 amino acids) and CspB (71 amino acids), respectively, Analyses of a cspC-lacZ transcriptional fusion and Northern (RNA) hybridization revealed that cspG is a low-temperature-responsive gene. Its low-temperature-inducible promoters were determined, and the results indicated that the cspG sequence is highly similar to both the cspA and cspB sequences not only in the coding regions but also in the 5'-upstream noncoding regions surrounding their own promoters.

  AMER SOC MICROBIOLOGY, May 1996, JOURNAL OF BACTERIOLOGY, 178(10) (10), 2994 - 2997, English

  [Refereed]

  Scientific journal


• CLONING AND SEQUENCING OF A SYNECHOCOCCUS GENE ENCODING A PROTEIN VERY SIMILAR TO MAMMALIAN ALDEHYDE DEHYDROGENASES

  S KASHIWAGI, J IRIE, K KANAMARU, T MIZUNO

  We report cloning and sequencing of a gene encoding a putative aldehyde dehydrogenase in Synechococcus sp. PCC7942. It was found that this phototrophic microorganism has a protein very similar to mammalian class-3 aldehyde dehydrogenases. A mutant strain lacking this gene was hypersensitive in growth to an aromatic aldehyde.

  TAYLOR & FRANCIS LTD, Dec. 1994, BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 58(12) (12), 2299 - 2300, English

  [Refereed]

  Scientific journal


• A COPPER-TRANSPORTING P-TYPE ATPASE FOUND IN THE THYLAKOID MEMBRANE OF THE CYANOBACTERIUM SYNECHOCOCCUS SPECIES PCC7942

  K KANAMARU, S KASHIWAGI, T MIZUNO

  P-type ATPases constitute a large family of cation pumps that play crucial physiological roles in many organisms, including bacteria, plants and mammals. They are postulated to play important roles in a variety of environmental adaptation systems. Recently, we cloned two distinct putative P-type ATPase genes (pacS and pacL) from a photosynthetic cyanobacterium, Synechococcus species PCC7942. In this study, one of the gene products (named PacS) was found to possess a putative metal-binding motif (Gly-Met-X-Cys-X-X-Cys) in its N-terminal portion. Thus we supposed that this ATPase may function as a metal pump. Indeed, the results of Northern blotting analysis showed that pacS-mRNA specifically increases upon addition of copper or silver to the growth medium. The results of Western blotting analysis confirmed the view that PacS accumulates in copper-treated Synechococcus cells. Thus we concluded that the expression of PacS ATPase is regulated in response to the change in concentration of external metals, namely copper and silver. Consistent with this, an insertional inactivation mutant of pacS exhibited hypersensitivity in terms of growth to these potentially toxic metals. It was also revealed that PacS was mainly located in the thylakoid membrane, in which the photosynthetic reactions take place. This P-type ATPase in the thylakoid membrane is implicated as a copper-transporting system that may be involved in copper-homeostasis crucial to the photosynthetic thylakoid function.

  BLACKWELL SCIENCE LTD, Jul. 1994, MOLECULAR MICROBIOLOGY, 13(2) (2), 369 - 377, English

  [Refereed]

  Scientific journal


• THE CYANOBACTERIUM, SYNECHOCOCCUS SP PCC7942, POSSESSES 2 DISTINCT GENES ENCODING CATION-TRANSPORTING P-TYPE ATPASES

  K KANAMARU, S KASHIWAGI, T MIZUNO

  P-type (or E1 E2-type) ATPases comprise a large family of prokaryotic and eukaryotic proteins capable of transporting a variety of cations. and function in a wide variety of cellular processes. The present study was carried out to search for genes encoding P-type ATPases in the phototrophic cyanobacterium, Synechococcus sp. PCC7942. We succeeded in cloning two genes each encoding P-type ATPases from this bacterium. It was found that Synechococcus at least, two distinct P-type ATPases; one belongs to the family of typical prokaryotic P-type ATPases and the other markedly resembles eukaryotic P-type ATPases. An insertion mutant lacking either of these two ATPase-genes was constructed. The results showed that the growth of these mutants is hypersensitive to osmotic stress upon addition of NaCl or sorbitol to the medium.

  ELSEVIER SCIENCE BV, Sep. 1993, FEBS LETTERS, 330(1) (1), 99 - 104, English

  [Refereed]

  Scientific journal


• SIGNAL TRANSDUCTION BETWEEN THE 2 REGULATORY COMPONENTS INVOLVED IN THE REGULATION OF THE KDPABC OPERON IN ESCHERICHIA-COLI - PHOSPHORYLATION-DEPENDENT FUNCTIONING OF THE POSITIVE REGULATOR, KDPE

  K NAKASHIMA, A SUGIURA, K KANAMARU, T MIZUNO

  The proteins KdpD and KdpE are crucial to the osmotic regulation of the kdpABC operon that is responsible for the high-affinity K+ ion transport system in Escherichia coli. We demonstrated previously that the response regulator, KdpE, is capable of undergoing phosphorylation mediated by the sensory protein kinase, KdpD. In this study, we obtained biochemical evidence supporting the view that when KdpE is phosphorylated, it takes on an active form that exhibits relatively high affinity for the kdpABC promoter, which in turn results in activation of the kdpABC operon. It was also suggested that the central hydrophobic domain of KdpD, which is conceivably responsible for membrane anchoring of this protein, plays a role in the signalling mechanism underlying KdpE phosphorylation in response to hyperosmotic stress.

  BLACKWELL SCIENCE LTD, Jan. 1993, MOLECULAR MICROBIOLOGY, 7(1) (1), 109 - 116, English

  [Refereed]

  Scientific journal


• SIGNAL TRANSDUCTION AND OSMOREGULATION IN ESCHERICHIA-COLI - A NOVEL MUTANT OF THE POSITIVE REGULATOR, OMPR, THAT FUNCTIONS IN A PHOSPHORYLATION-INDEPENDENT MANNER

  K KANAMARU, T MIZUNO

  In Escherichia coli, expression of the outer membrane proteins, OmpF and OmpC, is regulated by the regulatory factors, EnvZ and OmpR, at the transcriptional level in response to the medium osmolarity. In this particular osmotic regulation, phosphorylation of OmpR at an aspartate residue (Asp-55) by EnvZ plays an important role. The previously isolated mutant, ompR55Q, with the amino acid replacement of Asp-55 to Gln, exhibits an OmpF- and OmpC- phenotype. In this study, we isolated a novel type of ompR mutant, in which the defect caused by the ompR55Q mutation is suppressed. The intragenic suppressor mutation we isolated results in the amino acid replacement of Tyr-102 to Cys in the N-terminal domain of OmpR, and exhibits an OmpF+ and OmpC+ phenotype in response to the medium osmolarity in an EnvZ-independent manner. It was revealed that this amino acid replacement in OmpR enhances the in vitro DNA-binding ability to the cognate DNAs. These results suggested that OmpR is capable of functioning in a phosphorylation-independent manner under certain in vivo conditions, and further suggested that an EnvZ-independent mechanism may also be involved in the osmotically regulated expression of ompF and ompC.

  JAPANESE BIOCHEMICAL SOC, Apr. 1992, JOURNAL OF BIOCHEMISTRY, 111(4) (4), 425 - 430, English

  [Refereed]

  Scientific journal


• OSMOREGULATORY EXPRESSION OF THE PORIN GENES IN ESCHERICHIA-COLI - EVIDENCE FOR SIGNAL TITRATION IN THE SIGNAL TRANSDUCTION THROUGH ENVZ-OMPR PHOSPHOTRANSFER

  K NAKASHIMA, K KANAMARU, H AIBA, T MIZUNO

  The OmpR protein of Escherichia coli is a positive regulator specific for the ompF and ompC genes. The function of OmpR is modulated through phosphotransfer signaling mediated by the kinase, EnvZ. We previously demonstrated that OmpR contains two functional domains, which are physically separable; one is responsible for the interaction with EnvZ, whereas the other participates in interactions with cognate promoter DNAs. In this study, these domains of OmpR were overproduced in wild-type cells harboring the endogenous intact ompR gene on their chromosome. It was found that when the N-terminal domain of OmpR, which contains the phosphorylation site, was overproduced, expression of the ompF and ompC genes was markedly inhibited, irrespective of the osmolarity of the growth medium. Based on our current model for the molecular mechanism underlying signal transduction through Envz-OmpR phosphotransfer (T. Mizuno and S. Mizushima, Mol. Microbiol. 4, (1990), 1077-1082), we provide evidence that this phenomenon is best interpreted by the concept of 'signal titration' in the phosphotransfer signaling pathway.

  WILEY-BLACKWELL, Jul. 1991, FEMS MICROBIOLOGY LETTERS, 82(1) (1), 43 - 47, English

  [Refereed]

  Scientific journal


• Signal transduction and osmoregulation in Escherichia coli: A novel type of mutation in the phosphorylation domain of the activator protein, OmpR, results in a defect in its phosphorylation-dependent DNA binding

  K. Nakashima, K. Kanamaru, H. Aiba, T. Mizuno

  The transcriptional factors, OmpR and EnvZ, are crucially involved in the osmotic regulation of ompF and ompC expression in Escherichia coli. The DNA binding ability of the positive regulator, OmpR, is modulated through its phosphorylation and dephosphorylation mediated by EnvZ in response to the medium osmolarity. In this study, two examples of a novel type of mutant ompR allele, ompR96A and ompR115S, whose phenotype is OmpF- OmpC- irrespective of the medium osmolarity, were characterized. These mutations result in amino acid conversions, Glu96 to Ala and Arg115 to Ser, respectively, within the phosphorylation domain of OmpR. Nevertheless, these mutant proteins were capable of undergoing phosphorylation and dephosphorylation normally, just like wild-type OmpR. However, the phosphorylation-dependent enhancement of their in vitro DNA binding ability was found to be severely affected. It was thus revealed that these mutant OmpR represent a novel type in terms of the mechanism of phosphorylation-dependent activation of the function of OmpR, i.e. those are normally phosphorylated but not activated to bind to the cognate promoter DNAs. In this respect, it was further suggested that OmpR oligomerization may be involved in the mechanism underlying the phosphorylation-dependent enhancement of the DNA binding ability of OmpR. The mutant proteins characterized in this study seem to be defective in this particular oligomerization process observed in vitro.

  1991, Journal of Biological Chemistry, 266(17) (17), 10775 - 10780, English

  [Refereed]

  Scientific journal


• TRANSMEMBRANE SIGNAL TRANSDUCTION AND OSMOREGULATION IN ESCHERICHIA-COLI .1. ANALYSIS BY SITE-DIRECTED MUTAGENESIS OF THE AMINO-ACID-RESIDUES INVOLVED IN PHOSPHOTRANSFER BETWEEN THE 2 REGULATORY COMPONENTS, ENVZ AND OMPR

  K KANAMARU, H AIBA, T MIZUNO

  Previously, the transfer of a phosphoryl group between the EnvZ and OmpR proteins, which are involved in expression of the ompF and ompC genes in response to the medium osmolarity, was demonstrated in vitro. In this study, the histidine (His) residue at position 243 of the EnvZ protein, and the aspartate (Asp) residues at positions 12 and 55 of the OmpR protein were changed, re

  JAPAN BIOCHEMICAL SOC, Sep. 1990, JOURNAL OF BIOCHEMISTRY, 108(3) (3), 483 - 487, English

  [Refereed]

  Scientific journal


• Signal transduction in bacteria.

  KANAMARU Kengo, MIZUNO Takeshi

  Lead, Japan Society for Bioscience, Biotechnology, and Agrochemistry, 1990, KAGAKU TO SEIBUTSU, 28(8) (8), 498 - 505, Japanese

  [Invited]

  Scientific journal


• SIGNAL TRANSDUCTION AND OSMOREGULATION IN ESCHERICHIA-COLI - A SINGLE AMINO-ACID CHANGE IN THE PROTEIN-KINASE, ENVZ, RESULTS IN LOSS OF ITS PHOSPHORYLATION AND DEPHOSPHORYLATION ABILITIES WITH RESPECT TO THE ACTIVATOR PROTEIN, OMPR

  K KANAMARU, H AIBA, S MIZUSHIMA, T MIZUNO

  The EnvZ protein is a bacterial protein kinase, which specifically phosphorylates the activator protein, OmpR, involved in expression of the ompF and ompC genes in Escherichia coli. The phosphotransfer between the EnvZ and OmpR proteins was postulated to be involved in the signal transduction in response to an environmental osmotic stimulus. In this study, we isolated a novel t

  AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC, Dec. 1989, JOURNAL OF BIOLOGICAL CHEMISTRY, 264(36) (36), 21633 - 21637, English

  [Refereed]

  Scientific journal

• Dynamism of chloroplast tRNA expression in Arabidopsis

  Mototsugu Kitamura, Kazuki Yanagida, Tomohide Uno, Hiroshi Yamagata, Kengo Kanamaru

  OXFORD UNIV PRESS, 2007, PLANT AND CELL PHYSIOLOGY, 48, S117 - S117, English

  Summary international conference


• Functional involvement of cGMP and NO in light regulation of gene expression of soybean flavonoid-biosynthetic enzymes

  Kenji Suita, Maiko Mitsui, Tomohide Uno, Kengo Kanamaru, Hiroshi Yamagata

  OXFORD UNIV PRESS, 2007, PLANT AND CELL PHYSIOLOGY, 48, S65 - S65, English

  Summary international conference


• Analysis of a knock-out mutant of an Arabidopsis T7 phage-type RNA polymerase, RpoTp (RpoT;3)

  Ryosaku Inagaki, Pyoyun Park, Michio Kanechi, Hirokazu Tsukaya, Kazuki Yanagida, Nozomu Sakurai, Hideyuki Suzuki, Daisuke Shibata, Tomohide Uno, Hiroshi Yamagata, Kengo Kanamaru

  OXFORD UNIV PRESS, 2007, PLANT AND CELL PHYSIOLOGY, 48, S84 - S84, English

  Summary international conference


• An Arabidopsis double mutant sig2sig6 exhibits albino phenotype.

  Y Ishizaki, K Ozono, C Takenaka, Y Tsunoyama, Y Nakahira, K Tanaka, K Kanamaru, M Hanaoka, T Shiina

  Plastids contain a eubacterial-type RNA polymerase (PEP) that is responsible for transcription of photosynthesis genes encoded by plastid genome. Analogous to the eubacterial enzyme, PEP is a multi-subunit enzyme composed of a catalytic core complex (a2bbb) and a sigma factor that confers transcription specificity. Arabidopsis contains six plastid sigma factors (AtSig1-AtSig6) encoded by nuclear genome, although the core subunits are encode by plastid genome. Recently, general (AtSig6) and specialized (AtSig2 and AtSig5) sigma factors in plastids have been characterized by analyses of T-DNA tag mutants. To learn more on the functions of AtSig2 and AtSig6, we have invented double mutant plants (sig2sig6). The double mutant exhibited albino cotyledons and true leaves, although the single mutants showed pale green phenotypes. Interestingly, transcripts of psbB operon were seriously defected in the double mutant. These results suggested that AtSig2 and AtSig6 might play a major role in chloroplast transcription

  OXFORD UNIV PRESS, 2006, PLANT AND CELL PHYSIOLOGY, 47, S187 - S187, English

  Summary international conference


• 葉緑体分化における2種の色素体転写装置のスイッチング機構

  華岡光正, 金丸研吾, 藤原誠, 高橋秀夫, 田中寛

  2004, 日本分子生物学会年会プログラム・講演要旨集, 27th

• バイオスティミュラントハンドブック 植物の生理活性プロセスから資材開発、適用事例まで

  Contributor, 第4章 遺伝子による環境ストレス制御メカニズム／第3節 5-アミノレブリン酸（5-ALA）による遺伝子発現と環境ストレス耐性メカニズム, エヌ・ティー・エス, Apr. 2022, ISBN: 9784860437695


• Advances in Photosynthesis and Respiration, Vol. 36 "Plastid Development in Leaves during Growth and Senescence" #10. Dynamic Features of the Plastid Genome and its Transcriptional Control in Plastid Development

  KANAMARU KENGO, Sugita Mamoru

  Joint work, Springer, May 2013, English, The major function of chloroplasts in green plants and algae is oxygenic photosynthesis. Further, chloroplasts manufacture essential metabolites and phytohormones. Because of the extensive interconnection with cellular metabolic and regulatory networks, development of the organelle and the organism are coordinately regulated. Conversion of proplastids to chloroplasts is associa

  Scholarly book


• "Adaptive Gene Regulations from Microorganisms to Organelles" #4 Function of plastidial T7 phage- types RNA polymerase in plants

  KANAMARU KENGO

  Joint work, Research Signpost, Kerala, India, Dec. 2008, English

  Scholarly book


• PNE: Activity of The Japanese Consortium for Arabidopsis Thaliana DNA array (JCAA)

  藤原 徹, 太田 啓之, KANAMARU KENGO

  Others, 共立出版社, Jan. 2002, Japanese, ポストゲノム時代をむかえ，高価なアレイ解析をかずさDNA研究所からのcDNAライブラリーの提供などの協力の下，全国の植物研究者でコンソーシアムを組織してマクロアレイを共同作成した活動について紹介。

  Scholarly book

• Expression and Physiological Functions of Two Transcription Factors ERF105 and ERF104 in Arabidopsis

  Jingyan Xu, Kyoka Sato, Tomohide Uno, Daiki Hayashi, Kengo Kanamaru

  第65回日本植物生理学会, Mar. 2024, English

  Poster presentation


• Characterization of an Arabidopsis double mutant, which defects both transcription and division systems of chloroplasts

  Kyoka Satou, Daiki Hayashi, Tomohide Uno, Kengo Kanamaru

  第65回日本植物生理学会年会, Mar. 2024, English

  Poster presentation


• Characterization of enzyme activity, multimerization, and gene expression of two ALADs in Arabidopsis

  Yuri Kanbayashi, Masashi Amano, Tomohide Uno, Kengo Kanamaru

  第62回日本植物生理学会年会, Mar. 2021, English

  Oral presentation


• Physiological effects and molecular mechanisms triggered by exogenous s-4-hydroxy-5-amino valeric acid (s-HAVA)

  Kaho Tsuruyama, Fumiya Endo, Kaito Hinokawa, Noritaka Aoki, Takanori Fujimoto, Shigeyuki Watanabe, Tomohide Uno, Kengo Kanamaru

  第62回日本植物生理学会年会, Mar. 2021, English

  Oral presentation


• Transcriptional regulation in response to 5-aminolevulinic acid in Arabidopsis

  Imamura Nao, Tsiruyama Kaho, Iwamura Sakura, Sakamoto Minori, Kuroda Shuji, Uno Tomohide, KANAMARU KENGO

  JSPP, Mar. 2019, Japanese, Nagoya University, Domestic conference

  Oral presentation


• Overexpression and characterization of Arabidopsis 5-aminolevulinic acid dehydratases in E. coli

  Kanbayashi Yuri, UNO TOMOHIDE, KANAMARU KENGO

  10th International Symposium of iBioK, Jan. 2019, English, Kobe University, International conference

  Poster presentation


• 5-aminolevulinic acid responsive genes related to  increasing stress tolerance in Arabidopsis

  Tsuruyama Kaho, IMamura Nao, UNO TOMOHIDE, KANAMARU KENGO

  10th International Symposium of iBioK, Jan. 2019, English, Kobe University, International conference

  Poster presentation


• シロイヌナズナにおけるALA類縁物質による病原菌抵抗性の誘導

  Sakamoto Minori, KANAMARU KENGO

  2018若手フロンティア, Dec. 2018, Japanese, Kobe University, Domestic conference

  Poster presentation


• 5-aminolevulinate up-regulated genes and their physiological effects in Arabidopsis

  KANAMARU KENGO, Tanaka Takahiko, Iwamura Sakura, Kuroda Shuji, Uno Tomohide

  第91回日本生化学会大会, Sep. 2018, Japanese, 京都国際会議場, Domestic conference

  Poster presentation


• Function of 5-aminolevulinate membrane transporters in Arabidopsis

  Sakamoto Minori, Nakakita Takuma, Kuroda Shuji, Tanaka Takahiko, Uno Tomohide, KANAMARU KENGO

  第91回日本生化学会大会, Sep. 2018, Japanese, 京都国際会議場, Domestic conference

  Poster presentation


• 昆虫の低分子量GTP結合蛋 白質Rabの機能解析

  UNO TOMOHIDE, 尾崎屋 悠祐, KANAMARU KENGO

  第62回日本応用動物昆虫学会大会, Mar. 2018, Japanese, 鹿児島大学郡元キャンパス, Domestic conference

  Oral presentation


• 4-ヒドロキシ-5-アミノ-バレリン酸 (HAVA) によるシロイヌナズナの遺伝子発現応答

  IWAMURA SAKURA, TANAKA TAKAHIKO, SAKAMOTO MINORI, KURODA SHUJI, SAITO MASARU, IKEDA TOMOAKI, WATANABE SHIGEYUKI, UNO TOMOHIDE, KANAMARU KENGO

  日本農芸化学会2018年度大会, Mar. 2018, Japanese, Meijo University, Domestic conference

  Oral presentation


• Genetical and phenotypic responses to 5-aminolevulinate (ALA) in Arabidopsis

  IWMURA SAKURA, TANAKA TAKAHIKO, KANAMARU KENGO

  9th International Symposium of iBioK, Feb. 2018, English, iBioK, Kobe University, International conference

  Poster presentation


• 植物における5-アミノレブリン酸関連トランスポーターの局在性と機能

  NAKAKITA TAKUMA, KONISHI MAHO, TERASHITA KAZUKI, YAMAGATA HIROSHI, UNO TOMOHIDE, KANAMARU KENGO

  ConBio2017, Dec. 2017, Japanese, 日本分子生物学会, 神戸ポートアイランド, Domestic conference

  Poster presentation


• シロイヌナズナにおける5-アミノレブリン酸応答性遺伝子の機能的解析と分子機構

  TANAKA TAKAHIKO, DUAN CHEN, KURODA SHUJI, YAMAGATA HIROSHI, UNO TOMOHIDE, KANAMARU KENGO

  ConBio2017, Dec. 2017, Japanese, 日本分子生物学会, 神戸ポートアイランド, Domestic conference

  Poster presentation


• 5-アミノレブリン酸近縁物質による 「負」の遺伝子発現制御

  IWAMURA SAKURA, TANAKA TAKAHIKO, SAKAMOTO MINORI, KURODA SHUJI, UNO TOMOHIDE, KANAMARU KENGO

  若手フロンティア研究会2017, Dec. 2017, Japanese, 神戸大学研究基盤センター, Domestic conference

  Poster presentation


• 5-アミノレブリン酸近縁物質による 「正」の遺伝子発現制御

  TANAKA TAKAHIKO, DUAN CHEN, KURODA SHUJI, UNO TOMOHIDE, KANAMARU KENGO

  若手フロンティア研究会2017, Dec. 2017, Japanese, Kobe University, Domestic conference

  Poster presentation


• 5-アミノレブリン酸(ALA)の膜輸送機構 〜変異株における細胞内ALA濃度と遺伝子発現への影響

  NAKAKITA TAKUMA, KURODA SHUJI, UNO TOMOHIDE, KANAMARU KENGO

  若手フロンティア研究会2017, Dec. 2017, Japanese, Kobe University, Domestic conference

  Poster presentation


• HAVAの植物における分子機能

  KANAMARU KENGO

  コスモ石油中央研究所セミナー, Feb. 2017, Japanese, コスモ石油中央研究所, Domestic conference

  Oral presentation


• ALT1 and ALT2, 5-aminolevulinic acid (ALA) membrane transporters, in Arabidopsis

  NAKAKITA TAKUMA, KURODA SHUJI, YAMAGATA HIROSHI, UNO TOMOHIDE, KANAMARU KENGO

  8th International Symposium of iBioK, Feb. 2017, English, iBioK, 神戸大学, International conference

  Poster presentation


• ALR1 and ALR2, 5-aminolevulinic acid (ALA)-responsive transcription factors, in Arabidopsis

  TANAKA TAKAHIKO, DUAN CHEN, KURODA SHUJI, YAMAGATA HIROSHI, UNO TOMOHIDE, KANAMARU KENGO

  8th International Symposium of iBioK, Feb. 2017, English, iBioK, 神戸大学, International conference

  Poster presentation


• Molecular mechanisms of 5-aminolevulinic acid (ALA)-induced stress tolerance in Arabidopsis

  TANAKA TAKAHIKO, NAKAKITA TAKUMA, DUAN CHEN, KURODA SHUJI, YAMAGATA HIROSHI, UNO TOMOHIDE, KANAMARU KENGO

  第39回日本分子生物学会年会, Dec. 2016, Japanese, 日本分子生物学会, パシフィコ横浜, Domestic conference

  Poster presentation


• Function of Arabidopsis 5-aminolevulinic acid transporter, ALT1, ALT2

  NAKAKITA TAKUMA, KONISHI MAHO, TERASHITA KAZUKI, YAMAGATA HIROSHI, UNO TOMOHIDE, KANAMARU KENGO

  第39回日本分子生物学会年会, Dec. 2016, Japanese, 日本分子生物学会, パシフィコ横浜, Domestic conference

  Poster presentation


• ALA-induced gene expression and stress tolerance in Arabidopsis

  KANAMARU KENGO, TANAKA TAKAHIKO, DUAN CHEN, SAITO MASARU, FUJIMOTO TAKANORI

  4th International ALA&Porphyrin Symposium, IAPS4, Dec. 2016, English, ALA and Porphyrin Research Society, Ryoujun Hall, Nagasaki University School of Medicine, International conference

  Oral presentation


• 5-アミノレブリン酸誘導性転写応答による植物のマルチストレス耐性向上のメカニズム

  TANAKA TAKAHIKO, DUAN CHEN, KURODA SHUJI, YAMAGATA HIROSHI, UNO TOMOHIDE, KANAMARU KENGO

  日本農芸化学会関西支部第497回例会, Dec. 2016, Japanese, 日本農芸化学会関西支部, 神戸大学, Domestic conference

  Oral presentation


• 5-アミノレブリン酸(ALA)の膜輸送機構 〜変異株における細胞内ALA濃度と遺伝子発現への影響

  NAKAKITA TAKUMA, KURODA SHUJI, KONISHI MAHO, TERASHITA KAZUKI, YAMAGATA HIROSHI, UNO TOMOHIDE, KANAMARU KENGO

  若手フロンティア研究会2016, Dec. 2016, Japanese, 神戸大学研究基盤センター, Domestic conference

  Poster presentation


• 5-アミノレブリン酸(ALA)の分子機能 〜ALA誘導性遺伝子による植物のストレス耐性増強

  TANAKA TAKAHIKO, DUAN CHEN, KURODA SHUJI, YAMAGATA HIROSHI, UNO TOMOHIDE, KANAMARU KENGO

  若手フロンティア研究会2016, Dec. 2016, Japanese, 神戸大学研究基盤センター, Domestic conference

  Poster presentation


• 植物5-アミノレブリン酸(ALA)膜トランスポーターの局在性および機能解析

  NAKAKITA TAKUMA, KURODA SHUJI, KONISHI MAHO, TERASHITA KAZUKI, UNO TOMOHIDE, YAMAGATA HIROSHI, KANAMARU KENGO

  第１１回トランスポーター研究会, Jul. 2016, Japanese, Kyoto University, 5-アミノレブリン酸（ALA）は、ヘムやクロロフィル等テトラピロール化合物生合成の共通前駆体である。植物では低濃度のALA投与で成長促進効果が、高濃度投与で光傷害誘導による枯死効果があることから、ALA含有肥料はすでに上市され，除草剤としても検討がされてきた。更に近年、中濃度のALA投与で様々な環境ストレスへの耐性が向上することがわかり、我々は新規植物活性化剤としての用途開発を視野にその分子機構について基盤研究を進めている。そのひとつがALAトランスポーターである。ALAトランスポーターの同定と機能解析は、植物アミノ酸輸送系の新たな一例となるだけでなく、ALAの生理効果を最大化して農業に活用する技術の開発基盤にもなる。 我々は動物ALAトランスポーターの情報や，推定される構造的・機能的特徴，局在性予測に基づき植物ALAトランスポーターの候補遺伝子, Domestic conference

  Poster presentation


• Transcriptional Regulation and Improvement of Stress Tolerance Induced by 5-Amino Levulinic Acid

  KANAMARU KENGO, Duan Chen, Tanaka Takahiko, Nakakita Takuma, Masaru Saito, Fijimoto Takanori, Uno Tomohide, Yamagata Hiroshi

  第57回日本植物生理学会, Mar. 2016, Japanese, 岩手大学, Domestic conference

  Poster presentation


• Possible 5-aminolevulinic acid (ALA) transporters (ALT) in Arabidopsis

  Nakakita Takuma, Kuroda Shuji, Konishi Maho, Miyake Michiru, Terashita Kazuki, Saito Masaru, Fujimoto Takanori, Uno Tomohide, Yamagata Hiroshi, KANAMARU KENGO

  The 7th International Symposium of Innovative BioProduction Kobe (iBioK), Jan. 2016, English, Kobe University, International conference

  Poster presentation


• 5-aminolevulinic acid (ALA)-upregulated genes (ALU) and ALA-induced multi-stress tolerance in Arabidopsis

  Tanaka Takahiko, Duan Chen, Miyake Michiru, Saito Masaru, Fujimoto Takanori, Uno Tomohide, Yamagata Hiroshi, KANAMARU KENGO

  The 7th International Symposium of Innovative BioProduction Kobe (iBioK), Jan. 2016, English, Kobe University, International conference

  Poster presentation


• 5-aminolevulinic acid (ALA)-responsive transcription factors (ALR) and regulatory gene expression in Arabidopsis

  Duan Chen, Kuroda Shuji, Nakajima Haruka, Nomura Hironari, Saito Masaru, Fujimoto Takanori, Uno Tomohide, Yamagata Hiroshi, KANAMARU KENGO

  The 7th International Symposium of Innovative BioProduction Kobe (iBioK), Jan. 2016, English, Kobe University, International conference

  Poster presentation


• 植物をストレス耐性化する5-アミノレブリン酸の作用機構

  Duan Chen, Uno Tomohide, Yamagata Hiroshi, KANAMARU KENGO

  若手フロンティア2015, Dec. 2015, Japanese, Kobe University, Domestic conference

  Poster presentation


• 稲わらのバイオリファイナリーに向けた特性の解析と多様性の評価

  GOUDA TAKASHI, TERANUMA HIROSHI, SUEHIRO MIKI, KANAMARU KENGO, KAWAGUCHI HIDEO, OGINO CHIAKI, KONDO AKIHIKO, YAMAZAKI MASANORI

  神戸大学若手フロンティア2015, Dec. 2015, Japanese, 神戸大学, Domestic conference

  Poster presentation


• バイオリファイナリー利用に向けた稲わらの希硫酸前処理後グルコース含量の自然変異

  GODA TAKASHI, TERAMURA HIROSHI, SUEHIRO MIKI, KANAMARU KENGO, KAWAGUCHI HIDEO, OGINO CHIAKI, KONDO AKIHIKO, YAMAZAKI MASANORI

  日本農芸化学会関西支部第492例会, Dec. 2015, Japanese, 神戸大学, Domestic conference

  Oral presentation


• Molecular mechanism of 5-aminolevulinic acid (ALA)-dependent gene expression and increase of environmental stress tolerance in plant

  Duan Chen, Tanaka Takahiko, Nakakita Takuma, Konishi Maho, Masaru Saito, Fijimoto Takanori, Uno Tomohide, Yamagata Hiroshi, KANAMARU KENGO

  第38回日本分子生物学会年会, Dec. 2015, Japanese, 神戸ポートアイランド, Domestic conference

  Oral presentation


• Molecular mechanism of 5-aminolevulinic acid (ALA)-dependent gene expression and increase of environmental stress tolerance in plant

  Duan Chen, Tanaka Takahiko, Nakakita Takuma, Konishi Maho, Masaru Saito, Fijimoto Takanori, Uno Tomohide, Yamagata Hiroshi, KANAMARU KENGO

  第38回日本分子生物学会年会, Dec. 2015, Japanese, 神戸ポートアイランド, 植物バイオマスの増産は，近年バイオリファイナリー産業の原料としての重要性が高まっており，植物の潜在的な生命力や環境適応力を最大限引き出す植物活性化物質の活用はその一法として有望である。 そこで我々は5-アミノレブリン酸（ALA）の生理活性に注目した。ALAはほぼ全ての生物が有するテトラピロール合成経路の中間基質で，合成されたヘムやクロロフィルは種々の酸化反応酵素や光合成集光色素の活性中心をなす。一方，植物種子や幼苗をALAで前処理しておくと塩や低温ストレスなど様々な環境ストレスへの耐性が長期間高まる現象が観察されている。この分子機構を解明すれば，ALAを肥料成分として以外に，塩害土壌などへの可耕地拡大，気象変動等による生育不良の改善など植物活性化物質として植物バイオマス増産に活用できる。 我々はまずトマトのストレス応答性転写因子からALA誘導性も, Domestic conference

  Poster presentation


• 青色光/UV-AシグナルによるダイズELIP遺伝子の転写調節機構

  Akihiro Hohjo, Satoru Kuwamoto, UNO TOMOHIDE, KANAMARU KENGO, YAMAGATA HIROSHI

  日本農芸化学会2015年度大会, Mar. 2015, Japanese, 岡山大学, Domestic conference

  Oral presentation


• ヌクレオシド三リン酸によるACC酸化酵素の活性化機構の解析

  Hiroki Iio, Takuma Maekawa, UNO TOMOHIDE, KANAMARU KENGO, YAMAGATA HIROSHI

  日本農芸化学会2015年度大会, Mar. 2015, Japanese, 岡山大学, Domestic conference

  Oral presentation


• ククミシン遺伝子の果実特異的発現機構 -転写調節と形質転換トマトにおける発現解析-

  Okada Kaori, Miki Kohno, ATSUKO FURUKAWA, AIRI IEFUJI, UNO TOMOHIDE, KANAMARU KENGO, YAMAGATA HIROSHI

  日本農芸化学会2015年度大会, Mar. 2015, Japanese, Domestic conference

  Oral presentation


• AtNAS1プロモーター中のNO応答性シスエレメントの解析

  Takuya Uchida, Tatsuya Ohara, Shunsuke Yano, Moeko Doi, UNO TOMOHIDE, KANAMARU KENGO, YAMAGATA HIROSHI

  日本農芸化学会2015年度大会, Mar. 2015, Japanese, 岡山大学, Domestic conference

  Oral presentation


• 5-アミノレブリン酸(ALA)による植物遺伝子発現調節と環境ストレス耐性向上の分子機構

  Duan Chen, Tanaka Takahiko, Nakakita Takuma, Konishi Maho, Saitou Masaru, Fujimoto Hisanori, Uno Tomohide, Yamagata Hiroshi, KANAMARU KENGO

  BMB2015, Mar. 2015, Japanese, 神戸国際展示場, 植物バイオマスの増産は，近年バイオリファイナリー産業の原料としての重要性が高まっており，植物の潜在的な生命力や環境適応力を最大限引き出す植物活性化物質の活用はその一法として有望である。 そこで我々は5-アミノレブリン酸（ALA）の生理活性に注目した。ALAはほぼ全ての生物が有するテトラピロール合成経路の中間基質で，合成されたヘムやクロロフィルは種々の酸化反応酵素や光合成集光色素の活性中心をなす。一方，植物種子や幼苗をALAで前処理しておくと塩や低温ストレスなど様々な環境ストレスへの耐性が長期間高まる現象が観察されている。この分子機構を解明すれば，ALAを肥料成分として以外に，塩害土壌などへの可耕地拡大，気象変動等による生育不良の改善など植物活性化物質として植物バイオマス増産に活用できる。 我々はまずトマトのストレス応答性転写因子からALA誘導性も, Domestic conference

  Poster presentation


• 3P-083 Development of 5-aminolevulinic acid production using engineered S. cerevisiae

  Hara Kiyotaka, Saito Masaru, Morikawa Kana, Kato (Ishii) Hiroko, Nomura Hironari, Fujimoto Takanori, Kanamaru Kengo, Kondo Akihiko

  日本生物工学会大会講演要旨集, 2015, Japanese, 日本生物工学会


• 青色光/UV-AによるダイズELIP遺伝子の転写調節機構

  Akihiro Hohjo, Satoru Kuwamoto, UNO TOMOHIDE, KANAMARU KENGO, YAMAGATA HIROSHI

  神戸大学研究基盤センター若手フロンティア研究会, Dec. 2014, Japanese, ダイズELIP（early light-inducible protein）遺伝子（GmELIP）の発現は青色光/UV-A照射により特異的に誘導される。GmELIPプロモーター中のGT-1-like boxとG boxの2つのシスエレメントが青色光/UV-A応答に必要かつ十分な制御配列である。転写因子GmGT-1がこれら両シスエレメントに結合することを既に報告したがG-box配列はbZIP型転写因子の標的配列でもある。そこで、今回は青色光/UV-AによるGmELIPの発現誘導におけるbZIP型転写因子の機能を解析した。 ダイズ光独立栄養培養細胞（SB-P細胞）中のGmELIPのUV-Aによる発現誘導はシクロヘキシミドによって強く阻害されたことから、この過程には新たなタンパク質合成が必要であることが示唆された。DNAマイクロアレイ分析による網羅的解析に, Domestic conference

  Poster presentation


• ヌクレオシド三リン酸によるACC酸化酵素の活性化機構の解析

  HIROKI IIO, TAKUMA MAEKAWA, UNO TOMOHIDE, KANAMARU KENGO, YAMAGATA HIROSHI

  神戸大学研究基盤センター若手フロンティア研究会, Dec. 2014, Japanese, 神戸大学会館, エチレンは、花芽の形成や果実の追熟、器官脱離、老化、感染防御など様々な生理現象を制御している重要な植物ホルモンであり、メチオニンからS-アデノシルメチオニン、1-アミノシクロプロパン-1- カルボン酸（ACC）を経て合成される。ACC酸化酵素（ACO）はACCを酸化しエチレンに変換する最後の反応を触媒するが、その特性には不明な点が多い。本研究では、シロイヌナズナのAtACO2とトマトのLeACO1について新たな酵素学的特性を見出した。まず大腸菌で発現・精製したACOがアルカリ性で可逆的に失活することを明らかにした。次に、AtACO2の活性は12 mMヌクレオシド三リン酸（NTP）により約5倍に増加するのに対し、LeACO1は逆に約1/2に減少することを見出した。また、NTPのACCや各補因子に対するKm、Vmaxへの影響を調べた。一般的にNTPによる, Domestic conference

  Poster presentation


• ククミシン遺伝子の果実特異的発現機構

  Kaori Okada, Miki Kohno, Atsuko Furukawa, Airi Iefuji, UNO TOMOHIDE, KANAMARU KENGO, YAMAGATA HIROSHI

  神戸大学研究基盤センター若手フロンティア研究会, Dec. 2014, Japanese, 神戸大学会館, メロン果実のセリンプロテアーゼ、ククミシンは果実の全可溶性タンパク質の10％以上を占める主要タンパク質で、受粉後10日前後の若い果実の果芯部でのみ発現する。ククミシンプロモーターの果実特異的発現能はトマトにおいても機能することを報告した。また、ククミシン遺伝子のプロモーター中に存在する、G-boxを含む20塩基の配列がククミシンの果実特異的な発現を制御するエンハンサーとして機能することが明らかにされ、その配列に結合する転写因子候補として、CmbZIP1とCmbZIP2が単離された。また、CmbZIP1とCmbZIP2自体が果実特異的に発現することも報告された。本研究では、CmbZIP1とCmbZIP2の果実内動態とDNA結合特性の解明を目的とした。受粉後10-15 日のメロン果実の核タンパク質中に上記のククミシンエンハンサーに結合するDNA結合タンパ, Domestic conference

  Poster presentation


• AtNAS1プロモーター中のNO応答性シスエレメントの解析

  TAKUYA UCHIDA, TATSUYA OHARA, SHUNSUKE YANO, MOEKO DOI, UNO TOMOHIDE, KANAMARU KENGO, YAMAGATA HIROSHI

  神戸大学研究基盤センター若手フロンティア研究会, Dec. 2014, Japanese, 神戸大学会館, 一酸化窒素（NO）は植物において根の伸長や鉄の取り込み、気孔の閉鎖、植物ホルモンのシグナル伝達、病原菌感染や非生物的ストレスに対する適応応答等の様々な生理的過程に関与する重要なシグナル伝達分子である。動物に比べ、植物におけるNOシグナル伝達や遺伝子発現調節機構には不明な点が多い。本研究では、NOによる植物の遺伝子発現調節機構を転写レベルで明らかにすることを目的とした。シロイヌナズナの光独立栄養培養細胞（T87細胞）を用いたDNAマイクロアレイ解析により、NOによって発現が強く誘導される遺伝子としてニコチアナミン合成酵素遺伝子1（AtNAS1）が見出された。AtNAS1プロモーターの部分配列とGUSレポーター遺伝子の融合遺伝子をT87細胞のプロトプラストにPEG法で導入し、100 µM ニトロプルシドナトリウム（SNP、NO発生剤）で3時間処理後、GU, Domestic conference

  Poster presentation


• Transcriptional control of fruit-secific expression of melon cucumisin

  Okada Kaori, Miki Kohno, Tomohide Uno, KANAMARU KENGO, YAMAGATA HIROSHI

  Plant Biology 2014, Jul. 2014, English, アメリカ植物生物学会, アメリカ・オレゴン州ポートランド・コンベンションセンター, Cucumisin is a subtilisin-like serine protease found in melon fruits. We reported the primary structure and characterization of cucumisin (1, 2). Cucumisin is synthesized in the central parts of young fruits and secreted into the juice; it comprises more than 15% of the total juice protein. A 20 bp enhancer element in the cucumisin promoter is responsible for the fruit-specific, International conference

  Poster presentation


• シロイヌナズナRPOTmpの葉緑体光防護における役割

  KANAMARU KENGO, 山口 泰広, 李 棟梁, 一林 久雄, 野村 裕也, 宇野 知秀, 山形 裕士

  第55回日本植物生理学会年会, Mar. 2014, Japanese, 富山大学, Domestic conference

  Oral presentation


• カイコ脳における低分子量GTP結合蛋白質(Rab)の免疫組織学的解析

  YAMAGATA HIROSHI, 古谷 昌之, 坂元 一樹, KANAMARU KENGO, TAKEDA MAKIO, 溝口 明, UNO TOMOHIDE

  平成26年度 蚕糸・昆虫機能利用学術講演会, Mar. 2014, Japanese, Domestic conference

  Oral presentation


• 葉緑体・ミトコンドリア両局在性RNAPの光ストレス低減機能

  山口 泰広, 李 棟梁, 一林 久雄, 野村 裕也, 宇野 知秀, 山形 裕士, KANAMARU KENGO

  2013神戸大学研究基盤センター若手フロンティア研究会, Dec. 2013, Japanese, 神戸大学, Domestic conference

  Poster presentation


• 青色光/UV-AによるダイズELIP遺伝子の発現制御機構

  Kuwamoto Satoru, Hamad Abu Zahra, Nogi Takasuke, Uno Tomohide, KANAMARU KENGO, YAMAGATA HIROSHI

  第36回日本分子生物学会年会, Dec. 2013, Japanese, 神戸国際会議場, Domestic conference

  Poster presentation


• 青色光/UV-AによるダイズELIP遺伝子の発現制御機構

  桑本 知, Hamad Abu Zahra, 野木 貴祐, UNO TOMOHIDE, KANAMARU KENGO, YAMAGATA HIROSHI

  神戸大学研究基盤センター若手フロンティア研究会, Dec. 2013, Japanese, Domestic conference

  Poster presentation


• メロン・ククミシン遺伝子プロモーターのトマトにおける発現解析

  Okada Kaori, Miki Kohno, Furukawa Atsuko, Hasegawa Sayako, Uno Tomohide, Kanamaru Kengo, YAMAGATA HIROSHI

  第36回日本分子生物学会年会, Dec. 2013, Japanese, 神戸国際会議場, Domestic conference

  Poster presentation


• メロン・ククミシン遺伝子プロモーターのトマトにおける発現解析

  YAMAGATA HIROSHI, 岡田 香, 河野 美貴, 古川 温子, 長谷川 清子, UNO TOMOHIDE, KANAMARU KENGO

  神戸大学研究基盤センター若手フロンティア研究会, Dec. 2013, Japanese, Domestic conference

  Poster presentation


• メロン・ククミシンの果実特異的発現を調節する転写因子CmbZIP1/2の特性解析

  Miki Kohno, Okada Kaori, Furukawa Atsuko, Uno Tomohide, Kanamaru Kengo, YAMAGATA HIROSHI

  第36回日本分子生物学会年会, Dec. 2013, Japanese, 神戸国際会議場, Domestic conference

  Poster presentation


• メロン・ククミシンの果実特異的発現を調節する転写因子CmbZIP1/2の特性解析

  YAMAGATA HIROSHI, 河野 美貴, 岡田 香, 古川 温子, UNO TOMOHIDE, KANAMARU KENGO

  神戸大学研究基盤センター若手フロンティア研究会, Dec. 2013, Japanese, Domestic conference

  Poster presentation


• シロイヌナズナRPOTmpの光ストレス応答における重要性

  山口 泰広, 李 棟梁, 宇野 知秀, 山形 裕士, KANAMARU KENGO

  第36回日本分子生物学会年会, Dec. 2013, Japanese, 神戸国際会議場, Domestic conference

  Poster presentation


• Analysis of cis-element responsible for cGMP in the promoter of soybean chalcone synthase gene

  Hamad Abu Zahra, Satoru Kuwamoto, Tomohide Uno, Kengo Kanamaru, YAMAGATA HIROSHI

  第36回日本分子生物学会年会, Dec. 2013, English, 神戸国際会議場, Domestic conference

  Poster presentation


• A cis-element responsible for cGMP in the promoter of the soybean chalcone synthase gene

  Hamad Abu Zahra, 桑本 知, UNO TOMOHIDE, KANAMARU KENGO, YAMAGATA HIROSHI

  神戸大学研究基盤センター若手フロンティア研究会, Dec. 2013, English, Domestic conference

  Poster presentation


• 大腸菌で発現したウナギシトクロムP450(CYP1A9，CYP1C1)の機能解

  和泉 智保, 梶 悟, KANAMARU KENGO, YAMAGATA HIROSHI, 上西 由翁, 板倉 隆夫, UNO TOMOHIDE

  平成25年度日本水産学会秋季大会, Sep. 2013, Japanese, 三重大学, Domestic conference

  Oral presentation


• Transcriptional control of fruit-specific expression of melon cucumisin.

  YAMAGATA HIROSHI, Miki Kono, Kaori Okada, Tomohide Uno, Kengo Kanamaru

  25th Congress of the Scandinavian Plant Physiology Society, Aug. 2013, English, Scandinavian Plant Physiology Society, Helsinger, Denmark, International conference

  Poster presentation


• 免疫組織化学を用いたカイコ低分子量GTP結合タンパク質(Rab)の機能解析

  YAMAGATA HIROSHI, 坂元 一樹, 磯山 侑里, KANAMARU KENGO, 竹田 真生夫, 溝口 明, UNO TOMOHIDE

  日本蚕糸学会第83回大会, Mar. 2013, Japanese, つくば, Domestic conference

  Oral presentation


• Involvement of Arabidopsis RPOTmp in chloroplast photoprotection

  KANAMARU KENGO, YAMAGUCHI YASUHIRO, Li D, ICHIBAYASHI HISAO, NOMURA HIRONARI, UNO TOMOHIDE, YAMAGATA HIROSHI

  第55回日本植物生理学会年会, Mar. 2013, Japanese, Toyama University, An Arabidopsis nuclear-encoded T7 phage-type RNA polymerase (NEP) named RPOTmp is a mitochondria/plastid dual targeting protein and fuction for the mitochondrial transcription of some respiratory chain complex genes, plastidic transcription of rrn operon during early development, and compensation of RPOTp function at some extent. We focused on the RPOTmp function for envirronme, Domestic conference

  Oral presentation


• Function of Arabidopsis RPOTmp under light stress condition

  Yasuhiro Yamaguchi, Li Dongliang, Tomohide Uno, Hiroshi Yamagata, Hironari Nomura, KANAMARU KENGO

  第54回日本植物生理学会年会, Mar. 2013, Japanese, 岡山大学, Domestic conference

  Poster presentation


• A study of the molecular function of 5-aminolevulinic acid for enhancement of salt-stress tolerance in higher plants

  Hironari Nomura, Haruka Nakajima, Jun Li, Shigeyuki Funada, Seiji Nishikawa, Tomohide Uno, Hiroshi Yamagata, KANAMARU KENGO

  第54回日本植物生理学会年会, Mar. 2013, Japanese, 岡山大学, Domestic conference

  Poster presentation


• 5-アミノレブリン酸による植物塩ストレス耐性向上と遺伝子発現応答

  金丸 研吾, 中島 晴香, 山口 泰広, 李 潤, 舩田 茂行, 宇野 知秀, 山形 裕士, 野村 裕也

  第85回日本生化学会大会, Dec. 2012, Japanese, 福岡国際会議場, Domestic conference

  Poster presentation


• 色素体局在性脂肪酸合成律速酵素の発現と機能

  小島 志織, 山本 俊佑, 能勢 琢也, 朴 杓允, 宇野 知秀, 山形 裕士, KANAMARU KENGO

  日本農芸化学会2011年度大会, Mar. 2011, Japanese, 京都女子大学, Domestic conference

  Oral presentation


• 植物アセチルCoAカルボキシラーゼの発現と機能の解析

  小島 志織, 山本 俊佑, 能勢 琢也, 宇野 知秀, 山形 裕士, KANAMARU KENGO

  2010神戸大学研究基盤センター若手フロンティア研究会, Dec. 2010, Japanese, 神戸大学, Domestic conference

  Poster presentation


• トマトにおける5-アミノレブリン酸の分子・生理機能

  中島 晴香, 小島 志織, 古閑 太郎, 都築 由起子, 舩田 茂行, 李 潤, 宇野 知秀, 山形 裕士, KANAMARU KENGO

  BMB2010（第33回日本分子生物学会年会 第83回日本生化学会大会）, Dec. 2010, Japanese, 神戸ポートアイランド, Domestic conference

  Poster presentation


• 色素体σ因子SIG6と発現相関する新規PPRの機能

  益 祐美子, 宇野 知秀, 山形 裕士, KANAMARU KENGO

  日本農芸化学会2010年度大会, Mar. 2010, Japanese, 東京大学, Domestic conference

  Oral presentation


• 葉緑体発達に必須なシロイヌナズナPPRの分子機能解析.

  山本 俊佑, 小島 志織, 能勢 琢也, 稲垣 良作, 朴 杓允, 宇野 知秀, 山形 裕士, KANAMARU KENGO

  第32回日本分子生物学会年会, Dec. 2009, Japanese, パシフィコ横浜, Domestic conference

  Poster presentation


• 動植物におけるテトラピロール合成系化合物の生理活性と産業利用

  KANAMARU KENGO

  研究会ピロール系色素化合物の科学, Dec. 2009, Japanese, 神戸大学, Domestic conference

  [Invited]

  Invited oral presentation


• Molecular and physiological function of PPRTp, a pentatricopeptide repeat protein, in Arabidopsis (2)

  Shiori Kojima, Shunsuke Yamamoto, Takuya Nose, Tomohide Uno, Hiroshi Yamagata, KANAMARU KENGO

  WINPTech 2009, Dec. 2009, Japanese, 神戸大学, International conference

  Poster presentation


• Molecular and physiological function of PPRTp, a pentatricopeptide repeart protein, in Arabidopsis (1)

  Shunsuke Yamamoto, Shiori Kojima, Takuya Nose, Pyoyun Park, Tomohide Uno, Hiroshi Yamagata, KANAMARU KENGO

  WINPTech2009, Dec. 2009, Japanese, 神戸大学, International conference

  Poster presentation


• Molecular and physiological function of PPRS6a and PPRS6b, pentatricopeptide repeat proteins, in Arabidopsis

  Yumiko Masu, Tomohide Uno, Hiroshi Yamagata, KANAMARU KENGO

  WINPTech 2009, Dec. 2009, Japanese, 神戸大学, International conference

  Poster presentation


• 高等植物色素体の遺伝子発現と細胞機能の統御

  KANAMARU KENGO

  生化学若い研究者の会京都支部秋のセミナー, Nov. 2009, Japanese, 京都大学, Domestic conference

  [Invited]

  Invited oral presentation


• シロイヌナズナ色素体accD遺伝子のRNA編集に関与するPPRの多面的効果

  小島 志織, 山本 俊佑, 能勢 琢也, 朴 杓允, 宇野 知秀, 山形 裕士, KANAMARU KENGO

  日本生化学会 2009年度大会, Oct. 2009, Japanese, 神戸国際会議場, Domestic conference

  Oral presentation


• Intergenomic gene expression and primary material production in the ultimate symbiont, chloroplast

  KANAMARU KENGO

  The 5th International iBiok Workshop, Sep. 2009, English, 神戸大学, International conference

  Oral presentation


• シロイヌナズナにおける5-ALAと関連分子の生理・分子作用

  川畑 絵里, 宇野 知秀, 山形 裕士, KANAMARU KENGO

  日本農芸化学会2009年度大会, Mar. 2009, Japanese, 福岡国際会議場, Domestic conference

  Poster presentation


• Physiological and molecular functions of RPOTmp (RPOT;2) in Arabidopsis

  李 棟梁, 宇野 知秀, 山形 裕士, KANAMARU KENGO

  第50回日本植物生理学会年会, Mar. 2009, Japanese, 名古屋, Domestic conference

  Oral presentation


• Physiological and molecular functions of a RPOTp (RPOT;3)-related gene in Arabidopsis

  山本 俊佑, 能勢 琢也, 宇野 知秀, 山形 裕士, KANAMARU KENGO

  第50回日本植物生理学会年会, Mar. 2009, Japanese, 名古屋, Domestic conference

  Keynote oral presentation


• 色素体転写装置RPOTmp欠損変異株の生化学的解析

  李 棟梁, 宇野 知秀, 山形 裕士, KANAMARU KENGO

  2008神戸大学研究基盤センター若手フロンティア研究会, Dec. 2008, Japanese, 神戸大学, Domestic conference

  Poster presentation


• 色素体転写・翻訳装置と機能相関する因子の探索と解析

  益 祐美子, 宇野 知秀, 山形 裕士, KANAMARU KENGO

  2008神戸大学研究基盤センター若手フロンティア研究会, Dec. 2008, Japanese, 神戸大学, Domestic conference

  Poster presentation


• 色素体アセチルCoAカルボキシラーゼの発現と機能

  小島 志織, 山本 俊佑, 宇野 知秀, 山形 裕士, KANAMARU KENGO

  2008神戸大学研究基盤センター若手フロンティア研究会, Dec. 2008, Japanese, 神戸大学, Domestic conference

  Poster presentation


• 色素体T7ファージ型RNAポリメラーゼと高い発現相関性をもつPPRTpの機能

  山本 俊佑, 能勢 琢也, 宇野 知秀, 山形 裕士, KANAMARU KENGO

  日本農芸化学会関西支部第457回講演会, Dec. 2008, Japanese, 日本盛 酒蔵通り煉瓦館, Domestic conference

  Oral presentation


• 色素体T7ファージ型RNAポリメラーゼRpoTmpの機能解析

  李 棟梁, 一林 久雄, 宇野 知秀, 山形 裕士, KANAMARU KENGO

  第31回日本分子生物学会年会，第81回日本生化学会大会合同大会, Dec. 2008, Japanese, 神戸国際会議場, Domestic conference

  Oral presentation


• バクテリアも相談しながら大事業を成し遂げている

  KANAMARU KENGO

  先端膜工学研究推進機構. 第２回膜工学サロン, Dec. 2008, Japanese, 神戸大学, Domestic conference

  [Invited]

  Invited oral presentation


• 「生き物のつながりをゲノムから眺めたら？〜インターゲノミクスで拓くこれからの農学〜」植物は元バクテリアの葉緑体ともちつもたれつ

  KANAMARU KENGO

  平成20年度 神戸大学農学研究科公開講座, Nov. 2008, Japanese, 神戸大学, Domestic conference

  [Invited]

  Invited oral presentation


• テトラピロール化合物の新規生理機能とバイオプロダクション

  KANAMARU KENGO

  日本触媒学会「固体触媒とバイオリファイナリーの接点：現状と展望」, Nov. 2008, Japanese, 神戸大学, Domestic conference

  [Invited]

  Invited oral presentation


• Arabidopsis PPRTp is crucial for post-transcriptional regulation of plastidic accD gene.

  Shunsuke Yamamoto, Tomohide Uno, Hiroshi Yamagata, KANAMARU KENGO

  WINPTech 2008, Nov. 2008, Japanese, 神戸大学, International conference

  Poster presentation


• 葉緑体の遺伝子発現から細胞の機能統御へ

  KANAMARU KENGO

  玉川大学農学部セミナー, Oct. 2008, Japanese, 玉川大学, Domestic conference

  [Invited]

  Invited oral presentation


• 色素体ゲノムの転写と細胞機能の統御

  KANAMARU KENGO

  第19回関西光合成研究会, Sep. 2008, Japanese, 神戸学院大学, Domestic conference

  [Invited]

  Invited oral presentation


• Development and characterization of photo- autotrophic Arabidopsis T87 cells

  Li Dongliang, KANAMARU KENGO

  第19回関西光合成研究会, Sep. 2008, Japanese, 神戸学院大学, Domestic conference

  Poster presentation


• 葉緑体ゲノム転写制御からみた色素体分化

  華岡 光正, 田中 寛, KANAMARU KENGO

  特定領域研究「植物の環境応答戦略としてのオルガネラ分化」第四回若手ワークショップ, Aug. 2008, Japanese, 京都・聖護院御殿荘, Domestic conference

  Oral presentation


• 色素体転写装置と相関するＰＰＲの機能

  山本 俊佑, 能勢 琢也, KANAMARU KENGO

  特定領域研究「植物の環境応答戦略としてのオルガネラ分化」第四回若手ワークショップ, Aug. 2008, Japanese, 京都・聖護院御殿荘, Domestic conference

  Oral presentation


• 葉緑体一核の転写・翻訳系の相互作用からみた植物像

  KANAMARU KENGO

  兵庫バイオテクノロジー研究会“第72回定例会”, Mar. 2008, Japanese, 神戸大学農学部, Domestic conference

  [Invited]

  Invited oral presentation


• 緑色培養細胞の光独立栄養培養法の改良と細胞特性の解析

  李 棟梁, 宇野 知秀, 山形 裕士, KANAMARU KENGO

  2007神戸大学研究基盤センター若手フロンティア研究会, Dec. 2007, Japanese, 神戸大学, Domestic conference

  Poster presentation


• 植物色素体ヘテロ転写系の機能および細胞統御との関連性

  KANAMARU KENGO

  東京工業大学大学院生命理工学研究科公開セミナー, Dec. 2007, Japanese, 東京工業大学, Domestic conference

  [Invited]

  Invited oral presentation


• シロイヌナズナ5-ALAデヒドラターゼの生理機能および 酵素学的性質の解析

  天尾 雅, 宇野 知秀, 山形 裕士, KANAMARU KENGO

  2007神戸大学研究基盤センター若手フロンティア研究会, Dec. 2007, Japanese, 神戸大学, Domestic conference

  Poster presentation


• シロイヌナズナALAD1過剰発現体の解析

  天尾 雅, KANAMARU KENGO

  特定領域研究「植物の環境応答戦略としてのオルガネラ分化」第3回若手ワークショップ, Oct. 2007, Japanese, 瀬戸市, Domestic conference

  Oral presentation


• Chloroplast gene expression and its effect on whole plant function.

  KANAMARU KENGO

  Kobe University Frontier Technology Forum 2007 Satellite Workshop "Dynamics of Biological Networks in and around Plant Cells", Oct. 2007, English, 神戸大学, Domestic conference

  Oral presentation


• カイコGTP結合タンパク質（Rab）の機能解析

  森脇 翼, 中田 拓哉, 宇野 知秀, 金丸 研吾, 山形 裕士

  平成19年度蚕糸・昆虫利用学術講演会, Apr. 2007, Japanese, Domestic conference

  Oral presentation


• 葉緑体tRNAは動的に発現調節されている

  北村 元嗣, 柳田 一樹, 宇野 知秀, 山形 裕士, 金丸 研吾

  日本植物生理学会2007年度年会, Mar. 2007, Japanese, 松山, Domestic conference

  Oral presentation


• 高等生物のP450大腸菌発現系を用いたバイオコンバージョンシステムとその応用

  岡本 聡太, 宇野 知秀, 増田 智子, 伊東 鎮, 桝井 孝一, 山本 高明, 佐々木 彩子, 中西 昭二, 小野 雄介, 堀本 智仁, 金丸 研吾, 山形 裕士, 今石 浩正

  日本薬学会2007年度会, Mar. 2007, Japanese, 富山, Domestic conference

  Oral presentation


• 光シグナル、cGMPおよびNOによるダイズフラボノイド合成系酵素遺伝子群の発現調節

  吹田 憲治, 三井 麻衣子, 宇野 知秀, 金丸 研吾, 山形 裕士

  日本植物生理学会2007年度年会, Mar. 2007, Japanese, 松山, Domestic conference

  Oral presentation


• シロイヌナズナT7ファージ型RNAポリメラーゼRpoTp（RpoT;3）欠損株の解析

  稲垣 良作, 朴 杓允, 金地 通生, 塚谷 裕一, 柳田 一樹, 櫻井 望, 鈴木 秀幸, 柴田 大輔, 宇野 知秀, 山形 裕士, 金丸 研吾

  日本植物生理学会2007年度年会, Mar. 2007, Japanese, Domestic conference

  Oral presentation


• シロイヌナズナgatABCホモローグ遺伝子の局在性と機能

  伊藤 滋一, 櫻井 望, 鈴木 秀幸, 柴田 大輔, 宇野 知秀, 山形 裕士, 金丸 研吾

  2007年度日本農芸化学会, Mar. 2007, Japanese, 東京, Domestic conference

  Oral presentation


• ククミシン遺伝子の果実特異的発現機構の解析と果実の形質転換への応用

  奥山 慎也, 宇野 知秀, 金丸 研吾, 山形 裕士

  2007年度日本農芸化学会, Mar. 2007, Japanese, Domestic conference

  Oral presentation


• ククミシンプロ配列の自己阻害活性に重要な領域

  中川 真隆, 宇野 知秀, 金丸 研吾, 山形 裕士

  2007年度日本農芸化学会, Mar. 2007, Japanese, 東京, Domestic conference

  Oral presentation


• Analysis of a knock-out mutant of an Arabidopsis T7 phage-type RNA polymerase, RpoTp (RpoT;3)

  Ryosaku Inagaki, Pyoyun Park, Michio Kanechi, Hirokazu Tsukaya, Kazuki Yanagida, Nozomu Sakurai, Hideyuki Suzuki, Daisuke Shibata, Tomohide Uno, Hiroshi Yamagata, Kengo Kanamaru

  PLANT AND CELL PHYSIOLOGY, 2007, English, OXFORD UNIV PRESS


• Dynamism of chloroplast tRNA expression in Arabidopsis

  Mototsugu Kitamura, Kazuki Yanagida, Tomohide Uno, Hiroshi Yamagata, Kengo Kanamaru

  PLANT AND CELL PHYSIOLOGY, 2007, English, OXFORD UNIV PRESS


• Functional involvement of cGMP and NO in light regulation of gene expression of soybean flavonoid-biosynthetic enzymes

  Kenji Suita, Maiko Mitsui, Tomohide Uno, Kengo Kanamaru, Hiroshi Yamagata

  PLANT AND CELL PHYSIOLOGY, 2007, English, OXFORD UNIV PRESS


• 葉緑体tRNAの動的な発現調節とその分子機構

  北村 元嗣, 柳田 一樹, 宇野 知秀, 山形 裕士, 金丸 研吾

  神大会館, Dec. 2006, Japanese, Domestic conference

  Poster presentation


• 高等植物tRNAトランスアミデーション酵素の機能解析

  伊藤 滋一, 宇野 知秀, 山形 裕士, 金丸 研吾

  神戸大学研究基盤センター2006若手フロンティア研究会, Dec. 2006, Japanese, 神大会館, Domestic conference

  Poster presentation


• ククミシンプロ配列の自己阻害に重要な領域

  中川 真隆, 上山 恵, 宇野 知秀, 金丸 研吾, 山形 裕士

  神戸大学研究基盤センター2006若手フロンティア研究会, Dec. 2006, Japanese, 神大会館, Domestic conference

  Poster presentation


• Regulation of gene expression of soybean flavonoid-biosynthetic enzymes by cGMP

  吹田 憲治, 三井 麻衣子, 宇野 知秀, 金丸 研吾, 山形 裕士

  Kobe University Frontier Technology Forum, Nov. 2006, English, 神戸大学瀧川記念学術交流会館, Domestic conference

  Poster presentation


• Molecular function of a T7 phage-type RNA polymerase in chloroplasts

  稲垣 良作, 伊藤 滋一, 一林 久雄, 柳田 一樹, 宇野 知秀, 山形 裕士, 金丸 研吾

  Kobe University Frontier Technology Forum, Nov. 2006, English, 神戸大学瀧川記念学術交流会館, Domestic conference

  Poster presentation


• Basis and application of expression of two 5-ALA related enzymes in plants

  天尾 雅, 岩木 亜樹, 清田 真希, 高原 絵美, 宇野 知秀, 山形 裕士, 金丸 研吾

  Basis and application of expression of two 5’-ALA related enzymes in plants, Nov. 2006, English, 神戸大学瀧川記念学術交流会館, Domestic conference

  Poster presentation


• Characterization of plastidic glutamyl-tRNA synthetase and ALA dehydratase in Arabidopsis.

  岩木 亜樹, 金地 通夫, 岡田 秀樹, 鈴木 秀幸, 柴田 大輔, 岩井 一弥, 鈴木 貴也, 宇野 知秀, 山形 裕士, 金丸 研吾

  11th IUPAC, Aug. 2006, English, 神戸, International conference

  Poster presentation


• Pro-region of cucumisin precursor acts as a potent inhibitor of cucumisin

  中川 真隆, 上山 恵, 宇野 知秀, 金丸 研吾, 山形 裕士

  20th IUBMB International Congress of Biochemistry and Molecular Biology, Jun. 2006, English, IUBMB, 京都国際会館, International conference

  Poster presentation


• Function of T7 Phage-Type RNA Polymerases in Arabidopsis (2) : RpoTmp (RpoT;2)

  Hisao Ichibayashi, 稲垣 良作, 朴 杓允, 金地 通生, 伊藤 滋一, 櫻井 望, 鈴木 秀幸, 柴田 大輔, 宇野 知秀, 山形 裕士, 金丸 研吾

  The 53rd NIBB Conference "Dynamic Organelles in Plant", Jun. 2006, English, Okazaki, International conference

  Poster presentation


• Function of T7 Phage-Type RNA Polymerases in Arabidopsis (1) : RpoTp (RpoT;3)

  Inagaki Ryosaku, P. Pak, H. Tsukaya, Michio Kanechi, S. Itoh, Kazuki Ynagida, Nozomu Sakurai, Hideyuki Suzuki, Daisuke Shibata, 宇野 知秀, 山形 裕士, 金丸 研吾

  The 53rd NIBB Conference "Dynamic Organelles in Plant, Jun. 2006, English, Okazaki, International conference

  Poster presentation


• Correlative and compensatory function of heterologous transcription systems in plastids

  金丸 研吾, 稲垣 良作, 一林 久雄, 宇野 知秀, 柴田 大輔, 山形 裕士

  20th IUBMB International Congress of Biochemistry and Molecular Biology, Jun. 2006, English, 京都, International conference

  Poster presentation


• Cloning of cDNAs for Rab proteins from the brain of Bombyx mori and phosphorylation of their proteins expressed in Escherichia coli.

  中田 拓哉, 中尾 敦史, 山形 裕士, 金丸 研吾, 宇野 知秀

  20th IUBMB International Congress of Biochemistry and Molecular Biology, Jun. 2006, English, 京都, International conference

  Poster presentation


• ダイズフラボノイド合成系酵素遺伝子の発現調節におけるcGMPとNOの機能相関

  Suita Kenji, Mitsui Maiko, Uno Tomohide, Kanamaru Kengo, Yamagata Hiroshi

  日本農芸化学会大会2006年度大会, Mar. 2006, Japanese, 京都, Domestic conference

  Poster presentation


• シロイヌナズナT7ファージ型RNAポリメラーゼRpoT;2の機能解析

  Kengo Kanamaru, Ichibayashi Hisao, Inagaki Ryosaku, Ito Shigekazu, Kanachi Michio, Sakurai Nozomu, Suzuki Hideyuki, Shibata Daisuke, Uno Tomohide, Yamagata Hiroshi

  日本植物生理学会2006年度年会, Mar. 2006, Japanese, つくば, Domestic conference

  Oral presentation


• ククミシンプロ配列の自己阻害活性

  Nakagawa Masataka, Ueyama Megumi, Uno Tomohide, Kanamaru Kengo, Yamagata Hiroshi

  日本農芸化学会大会2006年度大会, Mar. 2006, Japanese, 京都, Domestic conference

  Poster presentation


• NO及びcGMPによって発現が調節されるシロイヌナズナ遺伝子の網羅的解析

  Ohara Tatsuya, Kiyota Maki, Ito Shigekazu, Inagaki Ryosaku, Suita Kenji, Sakurai Nozomu, Suzuki Hideyuki, Shibata Daisuke, Uno Tomohide, Kanamaru Kengo, Yamagata Hiroshi

  日本農芸化学会2006年度大会, Mar. 2006, Japanese, 京都, Domestic conference

  Poster presentation


• An Arabidopsis double mutant sig2sig6 exhibits albino phenotype.

  Y Ishizaki, K Ozono, C Takenaka, Y Tsunoyama, Y Nakahira, K Tanaka, K Kanamaru, M Hanaoka, T Shiina

  PLANT AND CELL PHYSIOLOGY, 2006, English, OXFORD UNIV PRESS, Plastids contain a eubacterial-type RNA polymerase (PEP) that is responsible for transcription of photosynthesis genes encoded by plastid genome. Analogous to the eubacterial enzyme, PEP is a multi-subunit enzyme composed of a catalytic core complex (a2bbb) and a sigma factor that confers transcription specificity. Arabidopsis contains six plastid sigma factors (AtSig1-AtSig6) encoded by nuclear genome, although the core subunits are encode by plastid genome. Recently, general (AtSig6) and specialized (AtSig2 and AtSig5) sigma factors in plastids have been characterized by analyses of T-DNA tag mutants. To learn more on the functions of AtSig2 and AtSig6, we have invented double mutant plants (sig2sig6). The double mutant exhibited albino cotyledons and true leaves, although the single mutants showed pale green phenotypes. Interestingly, transcripts of psbB operon were seriously defected in the double mutant. These results suggested that AtSig2 and AtSig6 might play a major role in chloroplast transcription


• シロイヌナズナT7ファージ型RNAポリメラーゼRpoT;2（RpoTmp)の機能解析

  一林 久雄, 稲垣 良作, 伊藤 滋一, 金地 通生, 櫻井 望, 鈴木 秀幸, 柴田 大輔, 宇野 知秀, 山形 裕士, 金丸 研吾

  第28回日本分子生物学会 講演要旨集p.449(f), Dec. 2005, Japanese, 日本分子生物学会, 福岡, Domestic conference

  Oral presentation


• 葉緑体DNA分析によるケシ科植物の識別

  松下 亨, 中川 真隆, 吹田 憲治, 金丸 研吾, 高津 正久, 守安 正恭, 山形 裕士

  日本法科学技術学会第11回学術集会, Nov. 2005, Japanese, 日本法科学技術学会, 大阪, Domestic conference

  Oral presentation


• シロイヌナズナT87細胞の独立/混合条件における光応答の解析

  清田 真希, 関本 佳世子, 櫻井 望, 鈴木 秀幸, 柴田 大輔, 宇野 知秀, 山形 裕士, 金丸 研吾

  第78回日本生化学会大会 講演要旨集p.868(f), Oct. 2005, Japanese, 日本生化学会, 神戸, Domestic conference

  Oral presentation


• ダイズELIP遺伝子の青色光／UV-A応答性発現を調節するシスエレメント

  野木 貴祐, 横山 輝之, 宇野 和秀, 金丸 研吾, 山形 裕士

  第12回日本光生物学協会年会 要旨集p.59(f), Aug. 2005, Japanese, 日本光生物学協会, 京都, Domestic conference

  Oral presentation


• シロイヌナズナT87細胞の混合栄養培養と独立栄養培養における光応答の転写解析

  金丸 研吾, 清田 真希, 関本 佳世子, 櫻井 望, 鈴木 秀幸, 柴田 大輔, 宇野 知秀, 山形 裕士

  第23回日本植物分子細胞生物学会 p.193(f), Aug. 2005, Japanese, 日本植物分子細胞生物学会, 京都, Domestic conference

  Oral presentation


• ダイズフラボノイド合成系遺伝子のcGMP およびNO による発現調節

  吹田 憲治, 三井 麻衣子, 宇野 知秀, 金丸 研吾, 山形 裕士

  第52回日本生化学会近畿支部例会 要旨集p.38(f), May 2005, Japanese, 日本生化学会近畿支部, 神戸大学, Domestic conference

  Oral presentation


• カイコのメチル化DNA 結合タンパク質の分子特性

  野村 由佳, 岩永 陽介, 中尾 淳史, 中村 正彦, 田嶋 正二, 金丸 研吾, 山形 裕士, 宇野 知秀

  第52回日本生化学会近畿支部例会 要旨集p.37(f), May 2005, Japanese, 日本生化学会近畿支部, 神戸大学, Domestic conference

  Oral presentation


• 昆虫の脳に存在する蛋白質のＰＫＣによるリン酸化およびその機能解析

  中田 拓哉, 中尾 淳史, 金丸 研吾, 山形 裕士, 宇野 知秀

  第75回蚕糸昆虫機能学術講演会, Apr. 2005, Japanese, 日本蚕糸学会, 東京, Domestic conference

  Oral presentation


• 1原子酸素添加酵素チトクロームP450 を用いたバイオコンビケムへの応用

  増田 智子, 中尾 淳史, 宇野 知秀, 谷口 雄規, 山形 裕士, 金丸 研吾, 大野 清治, 今石 浩正

  第20回Combinatorial Chemistry研究会, Apr. 2005, Japanese, Combinatorial Chemistry研究会, 大阪, Domestic conference

  Oral presentation


• 果実特異的遺伝子発現を調節するエンハンサー因子のcDNA クローニングと特性解析

  東 礼奈, 柴本 憲幸, 宇野 知秀, 金丸 研吾, 山形 裕士

  日本農芸化学会大会2005年度大会 要旨集p.150, Mar. 2005, Japanese, 日本農芸化学会, 札幌, Domestic conference

  Oral presentation


• ダイズフラボノイド合成系酵素遺伝子のｃGMPおよび光による発現調節

  吹田 憲治, 三井 麻衣子, 宇野 知秀, 金丸 研吾, 山形 裕士

  日本農芸化学会大会2005年度大会 要旨集p.149(f), Mar. 2005, Japanese, 日本農芸化学会, 札幌, Domestic conference

  Oral presentation


• シロイヌナズナ葉緑体tRNA の発現ダイナミクス

  柳田 一樹, 宇野 知秀, 金丸 研吾, 山形 裕士

  日本農芸化学会大会2005年度大会 要旨集p.150(f), Mar. 2005, Japanese, 日本農芸化学会, 札幌, Domestic conference

  Oral presentation


• イネ種子二機能性酵素インヒビター（RASI）の大腸菌における発現およびα� アミラーゼ阻害特性

  出口 正揮, 宇野 知秀, 金丸 研吾, 山形 裕士

  日本農芸化学会大会2005年度大会 要旨集p.311(f), Mar. 2005, Japanese, 日本農芸化学会, 札幌, Domestic conference

  Oral presentation


• １原子酸素添加酵素チトクロームP450 を用いたバイオコンビケムへの応用

  増田 智子, 中尾 敦史, 宇野 知秀, 谷口 雄規, 山形 裕士, 金丸 研吾, 大野 清春, 今石 浩正

  第21回Combinatorial Chemistry 研究会, 2005, Japanese, Combinatorial Chemistry 研究会, 東京, Domestic conference

  Oral presentation


• シロイヌナズナ葉緑体のストレス応答シグマ因子SIG5の解析

  永島明知, 華岡光正, 藤原誠, 鹿内利治, 金丸研吾, 高橋秀夫, 田中寛

  日本植物生理学会年会要旨集, 2004


• 葉緑体分化における2種の色素体転写装置のスイッチング機構

  華岡 光正, 金丸 研吾, 藤原 誠, 高橋 秀夫, 田中 寛

  第27回日本分子生物学会年会, 2004, Japanese, 日本分子生物学会, 未記入, Domestic conference

  Oral presentation


• 高等植物葉緑体におけるｔRNA発現の重要性とそのダイナミクス

  柳田 一樹, 宇野 知秀, 山形 裕士, 金丸 研吾

  日本農芸化学会関西支部第437回講演会), 2004, Japanese, 日本農芸化学会関西支部, 未記入, Domestic conference

  Oral presentation


• ナノマシンが集積した葉緑体が拓く可能性

  金丸 研吾

  第3回JBA・一日神戸大学, 2004, Japanese, 未記入, 未記入, Domestic conference

  Oral presentation


• ダイズELIP遺伝子プロモーターの青色光/紫外光に対する光応答能

  横山 輝之, 宇野 知秀, 金丸 研吾, 山形 裕士

  日本農芸化学会2004年度大会, 2004, Japanese, 日本農芸化学会, 未記入, Domestic conference

  Oral presentation


• イネ種子二機能性酵素インヒビター（RASI)の大腸菌における発現およびα-アミラーゼ阻害特性

  出口 正輝, 宇野 知秀, 金丸 研吾, 山形 裕士

  第25回種子生理生化学研究会, 2004, Japanese, 未記入, 未記入, Domestic conference

  Oral presentation


• ダイズフラボノイド合成系酵素遺伝子のcGMP による発現調節

  吹田 憲治, 三井 麻衣子, 宇野 知秀, 金丸 研吾, 山形 裕士

  日本農芸化学会関西支部第432回講演会, Dec. 2003, Japanese, 日本農芸化学会関西支部, 神戸大学, Domestic conference

  Oral presentation


• Transcriptional regulation of plastid genes by interaction of eukaryotic RPL32-like protein with NEP RNA polymerase in Arabidopsis thaliana.

  HANAOKA M, KANAMARU Kengo, TANAKA K, TAKAHASHI H

  ISPMB 2003, International Society of Plant Molecular Biology, Jun. 2003, English, 未記入, Barcelona, Spain, International conference

  Oral presentation


• シロイヌナズナの葉緑体におけるT7ファージ型RNAポリメラーゼ(NEP)と相互作用する真核型リボソームL32-likeタンパク質の機能解析

  華岡光正, 金丸研吾, 高橋秀夫, 田中寛

  日本分子生物学会年会プログラム・講演要旨集, 2003


• 細菌型因子を駆使した核戦略による葉緑体支配

  金丸 研吾

  東京理科大セミナー, 2003, Japanese, 東京理科大学, 未記入, Domestic conference

  Oral presentation


• 高等植物葉緑体の発達と遺伝子発現

  金丸 研吾

  かずさDNA研究所セミナー, 2003, Japanese, かずさDNA研究所, かずさDNA研究所セミナー室, Domestic conference

  Oral presentation


• シロイヌナズナ「JCAA アレイの入手方法、使用状況」

  金丸 研吾

  シロイヌナズナDNAアレイワークショップ, 2003, Japanese, 未記入, 未記入, Domestic conference

  Oral presentation


• Small GTP binding proteins from the brain of Bombyx mori.

  NAKAO A, KATSURAUMA C, UNO Tomohide, KANAMARU Kengo, YAMAGATA Hiroshi

  Japan-Korea Joint Seminar on Sericultural Sciences and Insect Industry, 2003, English, 未記入, 未記入, International conference

  Oral presentation


• シロイヌナズナの葉緑体転写制御におけるsigBの役割

  華岡光正, 金丸研吾, 田中寛, 高橋秀夫

  日本植物生理学会年会要旨集, 2001


• THE ROLE OF SIGB IN TRANSCRIPTIONAL REGULATION OF ARABIDOPSIS THALIANA CHLOROPLASTS :

  HANAOKA Mitsumasa, KANAMARU Kengo, TANAKA Kan, TAKAHASHI Hideo

  Plant and cell physiology, 2001, English, Japanese Society of Plant Physiologists


• PLASTID-ENCODED GENE EXPRESSION DEPENDING ON A NUCLEAR-ENCODED σFACTOR FOR TRANSLATION BURST IN DEVELOPING CHLOROPLASTS :

  KANAMARU Kengo, TANAKA Kan, TAKAHASHI Hideo

  Plant and cell physiology, 2001, English, Japanese Society of Plant Physiologists


• Characterization of Nuclear-Encoded Plastid RNA Polymerase Sigma Factors in Arabidopsis thaliana :

  FUJIWARA Makoto, NAGASHIMA Akitomo, NAKAZATO Emi, KANAMARU Kengo, TANAKA Kan, TAKAHASHI Hideo

  Plant and cell physiology, 2000, English, Japanese Society of Plant Physiologists


• CHARACTERIZATION OF CHLOROPLAST RNA POLYMERASE SIGMA FACTOR GENES IN Arabidopsis thaliana

  FUJIWARA Makoto, KANAMARU Kengo, TANAKA Kan, TAKAHASHI Hideo

  Plant and cell physiology, Mar. 1999, English


• ARABIDOPSIS NITRATE REDUCTASE (NR) IS PHOSPHORYLATED AND BINDS TO 14-3-3 PROTEINS (GF14s)

  KANAMARU Kengo, SU Wenpei, CRAWFORD Nigel M.

  Plant and cell physiology, May 1998, English

• HAVAの生理効果と分子機構

  金丸研吾

  コスモ石油, 共同研究, 2016 - 2020, Principal investigator


• バイオプロダクション次世代農工連携拠点形成

  近藤昭彦, 荻野千秋, 芦田均, 大澤朗, 吉田健一, 金丸研吾, 他

  文科省・JST, 先端融合領域イノベーション創出拠点形成プログラム, 振興調整費, 神戸大学, 2008 - 2018, Coinvestigator


• Identification of an encourage factor released by a plant

  Kanamaru Kengo

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research, Grant-in-Aid for Challenging Exploratory Research, Kobe University, Apr. 2015 - Mar. 2017, Principal investigator

  Positive plant-plant interaction, named companion plants, is known as phenomena but not proved well in view of molecular mechanism. Scientific clarification of the phenomena in molecular level, even in a part, would contribute both maintenance of agro-environment and sustainable agriculture mediated by spread of companion planting, and we succeeded. That is, we found an unidentified compound (encourage factor) contained in or emitted from roots of a specific plant promotes various plant's growth, and the encourage factor transcriptionally induces some transcription factors and stress responsive genes leading the growth promotion. Furthermore, we succeeded to identify about 10 candidates of the encourage factor. We have applied a patent based on these data with a chemical company.


• カモミール抽出物の植物生育促進効果の解析

  金丸研吾

  カネカ, 共同研究, 2016 - 2017, Principal investigator


• 食糧生産支援

  金丸研吾

  カネカ, カネカ-神戸大学包括連携, 共同研究, 神戸大学, 2010 - 2015, Principal investigator


• 5-ALAによる遺伝子発現

  金丸研吾

  共同研究（コスモ石油）, 2014 - 2014, Principal investigator


• Search and clarification of a novel molecular function of 5-amonolevulinate

  KANAMARU Kengo

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Grant-in-Aid for Scientific Research (C), Kobe University, 2011 - 2013, Principal investigator

  5-aminolevulinic acid (ALA) is a primary metabolite which almost all living organism produce by theirselves. It's metabolic products, tetrapyrrole compounds, play critical roles as heme in blood and various oxidases, siroheme in N/S assimilation enzymes, chlorophyll in plant, and vitamin B12. Furthermore, external ALA-feeding confers increase of resistance to environmental stress to plants. In this study, we examined carefully the phenomena and tried to clarify the molecular mechanisms. We found that exogenous ALA contributes recovery of heme but not directly of chlorophyll and identified ALA-inducible transcription factors from tomato and Arabidopsis.


• 植物細胞内シグナル伝達物質としてのｃＧＭＰ／ＮＯの機能解析

  山形 裕士

  科学研究費補助金／基盤研究(C), 2009

  Competitive research funding


• Molecular function correlation and control mechanism centered on the plastid heterotranscription system

  金丸 研吾, 大橋 幸男

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research on Priority Areas, Grant-in-Aid for Scientific Research on Priority Areas, Kobe University, 2007 - 2008, Principal investigator

  <目的>色素体ヘテロ転写系を構成するPEP(複数のσ因子が存在)とNEP(すなわちRpoTpとRpoTmp)相互のあるいは制御因子を介した相関性を紐解き、核と色素体のゲノム間相互作用によるオルガネラ分化、細胞機能、形態形成の統御ネットワークを解明する。 <成果>モデル植物シロイヌナズナにおいてNEPと発現相関性が高いPPR(35アミノ酸の繰り返し構造をもつ一群のタンパク質で特異的RNA結合能が示唆されている)のひとつに焦点を当てた。そして葉緑体に局在するこのタンパク質が脂肪酸合成の初期段階を律速する酵素の1サブユニット発現における転写後調節(mRNA前駆体の特定C残基がU残基デアミネーションされる反応)に関わっていることを欠損変異株の解析から明らかにした。さらにこの欠損株は葉緑体の発達不全だけでなく、植物固体全体の形態形成にも重篤な欠損な欠損を示し、色素体機能が細胞統御に及ぶことを示した。 <意義>究極の共生体である色素体は少なくとも双子葉類において遺伝子発現レベルで脂肪酸合成系の根幹を今なお握っており、「もちつもたれつ」の関係があることを明らかにした。同時に植物での脂肪酸合成を人為的に制御、向上させる標的分子としてこのPPRは有望である。同じ遺伝子における同様の転写後調節現象はアブラナ科、マツ、マメ類においても知られており、これらの油糧系作物の脂肪酸合成促進を当該PPRもしくはそのホモローグを用いて達成可能であると考え、シロイヌナズナで実際にその効果が示唆されたため国際特許を申請した。


• Identification of phototransducer and analysis of signaling network in plants

  YAMAGATA Hiroshi, KANAMARU Kengo

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B), Grant-in-Aid for Scientific Research (B), Kobe University, 2004 - 2006, Coinvestigator

  1. Several specific mutants of Arabidopsis for phytochrome A-signaling were isolated and back-crossed for 3 generation. 2. The phenotype of AtGC1-mutant was characterized. There was no difference between the mutant and wild-type plant. 3. A protein (named as GIP1) that interacts with heterotrimeric G protein alpha subunit was isolated using yeast two-hybrid system, and the interaction between GIP1 and G alpha was characterized. 4. Several genes for enzymes involved in flavonoid biosynthetic pathway in soybean were induced by cGMP, NO and light illumination. The inductions of gene expression by NO were inhibited by LY83583, suggesting the involvement of guanylate-cyclase. 5. For the induction of gene expression of soybean early light-inducible protein (ELIP) by blue/UV-A light, the combination of two cis-element, GT-1 like box and G-box elements were necessary. Soybean GmGT-1 cDNA was cloned and characterized.


• ヘテロ転写系の相関制御による色素体分化プロセスと環境応答系の解明

  金丸 研吾

  科学研究費補助金／特定領域研究, 2005, Principal investigator

  Competitive research funding


• Analysis of the preiotropic effects of a nucleus-encoded σ factor during chloroplast development

  KANAMARU Kengo

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C), Grant-in-Aid for Scientific Research (C), The University of Tokyo, 2001 - 2002, Principal investigator

  A eubacterial-type RNA polymerase (PEP) in higher plants plays crucial roles in chloroplast development. Six regulatory sigma factor genes (SIG1 to SIG6) encoded in the nuclear DNA are found in Arabidpopsis, however, the definite gene specificity of each sigma factor is still unknown. We recently identified an Arabidopsis recessive pale-green mutant sig2-1 (formerly named abc1) in which T-DNA is inserted in SIG2 (formerly named sigB ; Atlg08540). This mutant could develop almost normal etioplasts under dark conditions but barely develop the small chloroplasts with poor thylakoid membranes and stacked lamellar under light conditions. In the sig2-1 mutant, both chlorophyll and photosynthetic and photosynthesis-related proteins were remarkably reduced. However, amounts of mRNAs for these plastid-encoded proteins were not altered. Further analyzes revealed that several plastid-encoded tRNAs including trnE-UUC were drastically reduced in the mutant. The plastid-encoded tRNA-Glu is indispensable for not only protein synthesis (translation) but also tetrapyrrole (chlorophyll and heme) biogenesis. We could assign -35 like and -10 like sequences to upstream of these SIG2-dependent tRNA genes, which are conserved in Arabidopsis, tobacco, spinach, rice and maize. On the other hand, transcripts of nucleus-encoded T7 phage-type RNA polymerase (NEP)-dependent genes were steadily accumulated in this mutant. These results indicate that SIG2 plays crucial roles in chloroplast development mediated by expression of some plastid-encoded tRNAs and negative regulation of NEP.


• Macroarray analysis of nuclear genes involved in chloroplast development

  金丸 研吾

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research on Priority Areas (C), Grant-in-Aid for Scientific Research on Priority Areas (C), The University of Tokyo, 2001 - 2001, Principal investigator

  <マクロアレイのケミルミでの検出・解析法の確立>ビオチン標識したcDNAプローブをハイブリさせたマクロアレイを化学発光させて蛍光イメージアナライザーでシグナルを取り込み、数値化して解析する手法を確立した。本研究代表らが設立したJCAAは、昨夏以降、高品質、低価格での委託作成の段取りを整え、第二期メンバーを公募して全国90研究グループを超えるコンソーシアムになった。本研究代表らによるDNAスポット量やデプローブ条件の最適化を済ませ、マクロアレイ355セットを5月迄に配分できる予定である。(蛋白質・核酸・酵素2002,Vol47,91-93)。 <データベース構築とアレイ解析結果の対比>JCAAアレイ上のESTクローン用のデータベース(名大鈴木孝征氏作成)を改良し、検索対象を全推定遺伝子に拡大、オルガネラ移行予測検索結果などを加えたDARTを全国の研究者に公開した。このDBによりJCAAマクロアレイ上には全遺伝子の4割、葉緑体移行が予測される蛋白質の半分以上の遺伝子が網羅されていることがわかった。そこで手始めに、光漂自剤の影響と葉緑体発達不全変異株についてアレイ解析した。 <国内外での成果の位置づけ>葉緑体関連遺伝子のアレイ解析によるグループ化において本研究は世界的にもパイオニア的取り組みである。特に定量性でマイクロアレイより優れたマクロアレイを汎用性の高いケミルミ法でシロイヌナズナでも解析できるようにした意義は大きい。誰でも利用できる国産cDNAアレイを有志活動で供給する試みは共同作成から委託作成へと拡大、発展している。これらにより本研究遂行に十分な数のマクロアレイが確保できた。さらに、残ったPCR産物を活用して効率的にサブアレイ作成ができるシステムを新年度から動かす予定で、自身を含め我が国におけるシロイヌナズナのマクロアレイ研究は今後急速に成果を挙げると期待される。


• Nuclear control and sequentiality in gene expression of chloroplast constituents in higher plants

  金丸 研吾

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Encouragement of Young Scientists (A), Grant-in-Aid for Encouragement of Young Scientists (A), The University of Tokyo, 1999 - 2000

  高等植物葉緑体は個有のDNAをもち120前後の遺伝子がコードされている.これらは真正細菌型のRNAポリメラーゼ(PEP)とTワファージ型のNEPによって転写されるが葉緑体への分化には特にPEPが必須である.このコア酵素は葉緑体DNAにコードされているが調節因子のシグマファクターが少くとも6つ核にコードされていることを見い出した.中でもsigBは重要でこの欠損によってpale greenになり葉緑体が小さく貧弱になることから、又sigB自身の発現が発芽のごく初期から起こることから更に解析中である.一方藍藻に起源をもつ葉緑体は転写の他に分裂についてもバクテリアのシステムを一部そのまま利用している.そのひとつがMinDのホモローグであり、後に移行したこの遺伝子の発現はもうひとつの因子FtsZと同様のパターンを示しつつ、機能的には逆の働きをもっていた.すなわち、その大量発現によって分裂は抑制され一細胞に1〜2個の巨大葉緑体ができた.又このタンパク質の局在は葉緑体の内外両方である可能性も示唆された.更に遺伝学的にすでに知られていたarc6変異は表現型がAtMinDlの大量発現株と非常に類似していたがFtsZの発現はほぼ正常でAcMinDlの発現が数倍になっていることも確認できた.


• Analysis of the synthesis and action of the diffusible signaling factor homoserine lactone in Escherichia coli

  金丸 研吾

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Encouragement of Young Scientists (A), Grant-in-Aid for Encouragement of Young Scientists (A), National Institute of Genetics, 1995 - 1995, Principal investigator

  グラム陰性細菌の一部で、動植物への浸入、感染の初期過程にみられる菌体濃度依存的転写誘導は、拡散性化学シグナルとLuxRファミリーによる細胞間情報伝達系によって制御されている。大腸菌からも、LuxRホモローグSdiAが細胞分裂装置遺伝子ftsQAZに特異的な正の転写因子として報告されている。我々はSdiAがftsQAZ以外の何らかの酵素の発現も制御しているのではないかと考えた。そしてSdiAの過剰発現によって細胞内に27kDaのタンパク質が顕著に蓄積することをみつけた。アミノ酸配列を決定したところ、このタンパク質は一次胆汁酸脱水素酵素7α-ヒドロキシステロイドデヒドロゲナーゼであることが明らかとなった。そこでこの酵素の構造遺伝子hdhAを小原ライブラリーよりクローン化し、hdhA-lacZフュージョンによる解析とノーザン解析を行った。その結果、hdhAの総転写量は対数増殖期から定常期への増殖相移行に伴って20〜30倍に増加し、しかもhdhAの転写誘導はSdiAだけでなく増殖定常期特異的σ因子(σ38)にも依存していることが明らかとなった。さらにプライマー伸長法により、両因子は互いに34bp離れたプロモーターから別々に転写を制御し、対数増殖期にはSdiA依存的な転写が増加し続け、定常期突入直前で急速に減少、かわってσ38依存性の転写が一過的に起こっていることが示唆された。SdiA自身の発現はσ38に全く依存しないが、rpoS欠失株では対数増殖期のSdiA依存的なhdhAの転写量が増加した。化学シグナルの存在証拠は得られなかったが、SdiAとσ38による転写の独立二重制御はftsQAZでも機能しており、細胞分裂装置と二次胆汁酸合成酵素の発現が、増殖相の移行、もしくは細胞濃度上昇によって同様に転写制御されていると思われ、腸内環境における適応応答系の一例であることが期待される。今後、転写活性化の分子機構と二重制御の生理的意義についてさらに解析を進めたい。


• Analysis of coupling mechanism of cell division and LPS synthesis in Escherichia coli

  金丸 研吾

  Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research Grant-in-Aid for Encouragement of Young Scientists (A), Grant-in-Aid for Encouragement of Young Scientists (A), National Institute of Genetics, 1994 - 1994, Principal investigator

  当研究室が保有する400株以上の温度感受性大腸菌分裂変異株のうち染色体27分にマップされる7株について解析した。その結果、変異は全てkdsA遺伝子を含むDNA領域で相補された。kdsAの相補鎖にも未知のORFが存在するが、部位特異的変異法によって、KdsAのアミノ酸を置換することなく相補鎖側のORFの5'末端にstopコドンを導入したDNA断片でも、温度感受性分裂異常を相補できたことから、変異株ではkdsAが変異していることが予想された。変異株のkdsAの塩基配列決定の結果、GA transition(変異剤にNTG)によって、たしかに284a.a.からなるKdsAタンパク質に各々以下の1アミノ酸置換を起こす変異が同定された。 14Ala→Thr、73Gly→Asp、118Leu→Phe、203Ala→Thr、227Ala→Val、234Ala→Thr kdsAは外膜の物理的安定性に重要なリポ多糖の構成因子KDOの前駆体KDO-8ーリン酸の合成酵素をコードしており、変異株では外膜タンパク質の合成パターンに変化が見られたこともあり、外膜の不安定化、膜タンパク質(分裂装置構成タンパク質?)の局在異常によって分裂阻害が引き起こされていることが予想される。これまでの細胞分裂研究は、分裂装置タンパク質の側から主に進められてきたが、これらkdsA変異を用いた解析から膜合成そのものと細胞分裂の共役関係が明らかになるかも知れず、今後の生化学的解析が期待される。

• グローバルサイエンスキャンパスROOTプログラム第３期実践ステージアドバイザー

  Advisor