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Structural and functional analysis and its application of the yeast novel N-asetylransferase that detoxifies a proline analogue

Research Project

Project/Area Number 15380076
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Applied biochemistry
Research InstitutionFukui Prefectural University

Principal Investigator

TAKAGI Hiroshi  Fukui Prefectural University, Department of Bioscience, Professor, 生物資源学部, 教授 (50275088)

Co-Investigator(Kenkyū-buntansha) ODA Junichi  Fukui Prefectural University, Department of Bioscience, Professor, 生物資源学部, 教授 (50027041)
HIBI Takao  Fukui Prefectural University, Department of Bioscience, Associate Professor, 生物資源学部, 助教授 (00285181)
TAKAHASHI Masakazu  Fukui Prefectural University, Department of Bioscience, Assistant Professor, 生物資源学部, 助手 (80315837)
Project Period (FY) 2003 – 2005
Project Status Completed (Fiscal Year 2005)
Budget Amount *help
¥14,900,000 (Direct Cost: ¥14,900,000)
Fiscal Year 2005: ¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2004: ¥4,300,000 (Direct Cost: ¥4,300,000)
Fiscal Year 2003: ¥7,000,000 (Direct Cost: ¥7,000,000)
KeywordsYeast / N-Acetyltrasnferase / Azetidine-2-carboxylic acid / Anti-oxidant enzyme / Oxidative stress / Reactive oxygen species / Freezing stress / Ethanol stress / プロリンアナログ耐性
Research Abstract

The MPR1 gene (sigma 1278b gene for proline-analogue resistance) of the budding yeast Saccharomyces cerevisiae, which is found in the Σ1278b strain, encodes a novel N-acetyltransferase (Mpr1) that detoxifies the proline analogue azetidine-2-carboxylate (AZC). In this study, we analyzed the structure and function of Mpr1 as follows.
1)We found that mpr1-disrupted cells were hypersensitive to oxidative stresses such as H_2O_2 and heat shock and contained increased levels of reactive oxygen species (ROS). In contrast, expression of MPR1 leads to a decrease in ROS level and an increase in cell viability after oxidative treatments. These results indicate that Mpr1 can reduce intracellular ROS levels. It was also found that put2-disrupted yeast cells lacking Δ^1-pyrroline-5-carboxylate (P5C) dehydrogenease increased ROS levels and put2-disrupted cells that expressed MPR1 had considerably lower ROS levels. Mpr1 acetylates P5C or its equilibrium compound glutamtate-γ-semialdehyde. These results … More suggest that P5C is toxic to yeast cells due to the formation of ROS and that Mpr1 regulates the ROS level under P5C-induced oxidative stress.
2) We found that yeast cells exhibited increased levels of ROS during freezing and thawing. Gene disruption and expression experiments indicated that Mpr1 protects yeast cells from freezing stress by reducing the intracellular levels of ROS. Similar results were obtained from ethanol stress. Similar results were obtained from ethanol stress. Gene disruption and expression experiments indicated that Mpr1 protect yeast cells from ethanol stress by reducing the intracellular ROS.
3) We investigated whether MPR1 would function similarly in bacterial and plant cells and if its expression could confer resistance to AZC. Our studies confirm that MPR1 can function in a similar fashion in Escherichia coli and tobacco as in yeast to detoxify the toxic AZC so could potentially be used as a new selectable marker for E.coli and plant transformation.
In summary, as a novel antioxidant mechanism, Mpr1 is supposed to reduce the intracellular oxidation level by acetylating the toxic metabolite(s) involved in ROS production, such as P5C/GSA. The proposed role of Mpr1 against oxidative stress could be promising for breeding novel stress-resistant yeast strains. Less

Report

(4 results)
  • 2005 Annual Research Report   Final Research Report Summary
  • 2004 Annual Research Report
  • 2003 Annual Research Report
  • Research Products

    (18 results)

All 2006 2005 2004 Other

All Journal Article (14 results) Book (2 results) Publications (2 results)

  • [Journal Article] 酵母に発見した「アセチル化酵素Mpr1」による新しい抗酸化メカニズム活性酸素種の生成に関わる物質を基質とした酸化ストレス防御システム?!2006

    • Author(s)
      高木 博史
    • Journal Title

      化学と生物 44

      Pages: 83-84

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Novel anti-oxidant mechanism by Mpr1 acetyltransferase found in yeast. (In Japanese)2006

    • Author(s)
      Hiroshi Takagi
    • Journal Title

      Kagaku and Seibutsu 44

      Pages: 83-84

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] 酵母に発見した「アセチル化酵素Mpr1」による新しい抗酸化メカニズム活性酸素種の生成に関わる物質を基質とした酸化ストレス防御システム2006

    • Author(s)
      高木 博史
    • Journal Title

      化学と生物 44

      Pages: 83-84

    • Related Report
      2005 Annual Research Report
  • [Journal Article] N-Acetyltransferase Mpr1 confers freeze tolerance on Saccharomyces cerevisiae by reducing reactive oxygen species2005

    • Author(s)
      Xiaoyi Du, Hiroshi Takagi
    • Journal Title

      Journal of Biochemistry 138

      Pages: 391-397

    • NAID

      10017347511

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Annual Research Report 2005 Final Research Report Summary
  • [Journal Article] N-Acetyltransferase Mpr1 confers freeze tolerance on Saccharomyces cerevisiae by reducing reactive oxygen species.2005

    • Author(s)
      Xiaoyi Du, Hiroshi Takagi
    • Journal Title

      Journal of Biochemistry 138

      Pages: 391-397

    • NAID

      10017347511

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Novel anti-oxidant mechanism by Mpr1 acetyltransferase found in yeast. (In Japanese)2005

    • Author(s)
      Hiroshi Takagi
    • Journal Title

      Report of Organization for Research Promotion of Health and Longevity (Fukui Prefectural University)

      Pages: 1-8

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Expression of a novel yeast gene that detoxifies the proline analog azetidine-2-carboxylate confers resistance during tobacco seed germination, callus and shoot formation2004

    • Author(s)
      Xing-Hai Zhang, Hiroshi Takagi, Jack.M.Widholm
    • Journal Title

      Plant Cell Reports 22

      Pages: 615-622

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Characterization of end4^+, a gene required for endocytosis in Schizosaccharomyces pombe2004

    • Author(s)
      Tomoko Iwaki, Naotaka Tanaka, Hiroshi Takagi, Yuko Giga-Hama, Kaoru Takegawa
    • Journal Title

      Yeast 21

      Pages: 876-881

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Role of the yeast novel acetyltransferase Mpr1 in oxidative stress : Regulation of oxygen reactive species caused by a toxic proline catabolism intermediate2004

    • Author(s)
      Michiyo Nomura, Hiroshi Takagi
    • Journal Title

      Proceedings of the National Academy of Sciences, U.S.A. 101

      Pages: 12616-12621

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Expression of a novel yeast gene that detoxifies the proline analog azetidine-2-carboxylate confers resistance during tobacco seed germination, callus and shoot formation.2004

    • Author(s)
      Xing-Hai Zhang, Hiroshi Takagi, Jack M.Widholm
    • Journal Title

      Plant Cell Reports 22

      Pages: 615-622

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Characterization of end4^+, a gene required for endocytosis in Schizosaccharomyces pombe.2004

    • Author(s)
      Tomoko Iwaki, Naotaka Tanaka, Hirohsi Takagi, Yuko Giga-Hama, Kaoru Takegawa
    • Journal Title

      Yeast 21

      Pages: 867-881

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Role of the yeast novel acetyltransferase Mpr1 in oxidative stress : Regulation of oxygen reactive species caused by a toxic proline catabolism intermediate.2004

    • Author(s)
      Michiyo Nomura, Hiroshi Takagi
    • Journal Title

      Proceedings of the National Academy of Sciences, U.S.A. 101

      Pages: 12616-12621

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Characterization of end4^+, a gene required for endocytosis in Schizosaccharomyces pombe2004

    • Author(s)
      T.Iwaki, N.Tanaka, H.Takagi, Y.Giga-Hama, K.Takegawa
    • Journal Title

      Yeast Vol.21 No.10

      Pages: 876-881

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Role of the yeast novel acetyltransferase Mpr1 in oxidative stress : Regulation of oxygen reactive species caused by a toxic proline catabolism intermediate2004

    • Author(s)
      M.Nomura, H.Takagi
    • Journal Title

      Proceedings of the National Academy of Sciences, U.S.A. Vol.101 No.34

      Pages: 12616-12621

    • Related Report
      2004 Annual Research Report
  • [Book] 健康長寿研究成果報告書 酵母に見いだした酵素Mpr1よる新しい抗酸化メカニズム2005

    • Author(s)
      高木 博史(分担執筆)
    • Publisher
      発行:福井県立大学、印刷:株式会社エクシート
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Book] 健康長寿研究成果報告書 酵母に見いだした酵素Mpr1による新しい抗酸化メカニズム2005

    • Author(s)
      高木 博史(分担執筆)
    • Publisher
      福井県立大学(印刷:株式会社エクシート)
    • Related Report
      2005 Annual Research Report
  • [Publications] Michiyo Nomura, Shigeru Nakamori, Hiroshi Takagi: "Characterization of novel aceryltransferases found in the budding and fission yeasts that detoxifies a proline analogue, azetidine-2-carboxylic acid"Journal of Biochemistry. Vol.133 No.1. 67-74 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Xing-Hai Zhang, H.Takagi, Jack.M.Widholm: "Expression of a novel yeast gene that detoxifies the proline analog azetidine-2-carboxylate confers resistance during tobacco seed germination, callus and shoot formation"Plant Cell Reports. Vol.22 No.8. 615-622 (2004)

    • Related Report
      2003 Annual Research Report

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Published: 2003-04-01   Modified: 2016-04-21  

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