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Proteome analysis on the regulation mechanism of antibiotic production

Research Project

Project/Area Number 10480167
Research Category

Grant-in-Aid for Scientific Research (B).

Allocation TypeSingle-year Grants
Section一般
Research Field Functional biochemistry
Research InstitutionOsaka University

Principal Investigator

NIHIRA Takuya  Department of Biotechnology, Graduate School of Engineering, Osaka University. Associate Professor, 大学院・工学研究科, 助教授 (70144441)

Co-Investigator(Kenkyū-buntansha) KINOSHIMA Hiroshi  Department of Biotechnology, Graduate School of Engineering, Osaka University. Assistant Professor, 大学院・工学研究科, 助手 (20294035)
YAMADA Yasuhiro  Department of Applied Biological Science, Faculty of Engineering Fukuyama University. Professor, 工学部, 教授 (00011891)
Project Period (FY) 1998 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥11,900,000 (Direct Cost: ¥11,900,000)
Fiscal Year 2000: ¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 1999: ¥4,000,000 (Direct Cost: ¥4,000,000)
Fiscal Year 1998: ¥5,100,000 (Direct Cost: ¥5,100,000)
KeywordsStreptomyces virginiae / autoregulator / Virginiae butanolide / 2D electrophoresis / antibiotic production / 2次元電気泳動
Research Abstract

In Streptomycetes, low-molecular-weight signal compounds called γ-butyrolactone autoregulators regulate the production of secondary metabolites, such as antibiotics and anticancer drugs.) From S.virginiae we have isolated virginiae butanolide (VB) a representative γ-butyrolactone autoregulator, and have clarified that the VB signal is transmitted via binding with a specific receptor protein, which finally to the production of secondary metabolites. Although many genes should participate in the signal transduction system, only a little knowledge is available on the pathway or the regulatory system until now.
In this study, we investigated the regulation mechanism of virginiamycin M and S (VM and VS) production in S.virginiae by proteome and transcriptome analysis.
To identify genes involved in the regulation, transcriptional patterns of several genes were compared by RT-PCR among wild-type strain, a disruptant of barA encoding a VB receptor and a disruptant of barX encoding a newly identified multi-functional regulator. It was demonstrated that transcription of genes encoding self-resistant machineries toward VS and VM were induced by VS and VM, respectively. Detailed regulatory mechanism by VB/VS/VM was investigated. Furthermore, novel genes under control of VB-BarA were identified. Proteome analysis by means of two dimensional electrophoresis resulted in the finding of several proteins showing different expression pattern between the wild type strain and the two mutant strains. Based on these results the whole regulatory mechanism of VS/VM production will be clarified in the near future.

Report

(4 results)
  • 2000 Annual Research Report   Final Research Report Summary
  • 1999 Annual Research Report
  • 1998 Annual Research Report
  • Research Products

    (26 results)

All Other

All Publications (26 results)

  • [Publications] Wises Namwat: "Identification of the varR gene as a transcriptional regulator of virginiamycin S resistance in Streptomycesvirginiae."J.Bacteriol.. 183(6). 2025-2031 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Ryu Kawachi: "Identification by gene deletion analysis of a regulator, VmsR, that controls virginiamycin biosynthesis in Streptomyces virginiae."J.Bacteriol.. 182(21). 6259-6263 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Ryu Kawachi: "Identification of an AfsA homologue (BarX) from Streptomyces virginiae as a pleiotropic regulator controlling autoregulator biosynthesis virginiamycin biosynthesis and virginiamycin resistance"Mol.Microbiol.. 36(2). 302-313 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Hiroshi Kinoshita: "Characterization of binding sequence for butyrolactone-autoregulator-receptors in Streptomycetes."J.Bacteriol.. 181(16). 5075-5080 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Chang-Kwon Lee: "Identification and in vivo functional analysis of a virginiamycin S resistance fene (varS) from Streptomyces virginiae."J.Bacteriol.. 181(10). 3293-3297 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Noriyasu Shikura: "Identification and characterization of 6-dehydro VB-A reductase from Streptomyces antibioticus"FEMS Microbiol.Lett.. 171. 183-189 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Namwat W.et al.: "Identification of the varR gene as a transcriptional regulator of virginiamycin S resistance in Streptomycesvirginiae."J.Bacteriol.. 183(6). 2025-2031 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kawachi R.et al.: "Identification by gene deletion analysis of a regulator, VmsR, that controls virginiamycin biosynthesis in Streptomyces virginiae."J.Bacteriol.. 182(21). 6259-6263 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kawachi R.et al.: "Identification of an AfsA homologue (BarX) from Streptomyces virginiae as a pleiotropic regulator controlling autoregulator biosynthesis virginiamycin biosynthesis and virginiamycin resistance"Mol.Microbiol.. 36(2). 302-313 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kinoshita H.et al.: "Characterization of binding sequence for butyrolactone-autoregulator-receptors in Streptomycetes."J.Bacteriol.. 181(16). 5075-5080 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Lee C.-K.et al.: "Identification and in vivo functional analysis of a virginiamycin S resistance fene (varS) from Streptomyces virginiae."J.Bacteriol.. 181(10). 3293-3297 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Shikura N.et al.: "Identification and characterization of 6-dehydro VB-A reductase from Streptomyces antibioticus"FEMS Microbiol.Lett.. 171. 183-189 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Wises Namwat: "Identification of the varR gene as a transcriptional regulator of virginiamycin S resistance in Streptomycesvirginiae."J. Bacteriol.. (In press).

    • Related Report
      2000 Annual Research Report
  • [Publications] Ryu Kawachi: "Identification by gene deletion analysis of a regulator, VmsR, that controls virginiamycin biosynthesis in Streptomyces virginiae."J.Bacteriol.. 182(21). 6259-6263 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Ryu Kawachi: "Identification of an AfsA homologue (BarX) from Streptomyces virginiae as a pleiotropic regulator controlling autoregulator biosynthesis virginiamycin biosynthesis and virginiamycin resistance"Mol.Microbiol.. 36(2). 302-313 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Hiroshi Kinoshita: "Characterization of binding sequence for butyrolactone-autoregulator-receptors in Streptomycetes."J.Bacteriol.. 181(16). 5075-5080 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] Chang-Kwon Lee: "Identification and in vivo functional analysis of a virginiamycin S resistance fene (varS) from Streptomyces virginiae."J.Bacteriol.. 181(10). 3293-3297 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] Noriyasu Shikura: "Identification and characterization of 6-dehydro VB-A reductase from Streptomyces antibioticus"FEMS Microbiol. Lett.. 171. 183-189 (1999)

    • Related Report
      2000 Annual Research Report
  • [Publications] Eriko Takano: "Purification and structural determination of SCB1, aγ-butyrolactone that elicits antibiotic production in Streptomyces coelicolor A3(2)"J. Biol. Chem.. (In Press).

    • Related Report
      1999 Annual Research Report
  • [Publications] Ryu Kawachi: "Identification of an AfsA homologue(BarX) from Streptomyces virginiae as a pleiotropic regulator controlling autoregulator biosynthesis, virginiamycin biosynthesis and virginiamycin resistance"Mol. Microbiol.. (In press).

    • Related Report
      1999 Annual Research Report
  • [Publications] Hiroshi Kinoshita: "Characterization of binding sequence for butyrolactone-autoregulator-receptors in Streptomycetes"J. Bacteriol.. 181(16). 5075-5080 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Shigeru Kitani: "In vitro analysis of the butyrolactone autoregulator receptor protein(FarA) of Streptomyces lavendulae FRI-5 reveals that FarA acts as a DNA-binding transcriptional regulator that controls its own synthesis"J. Bacteriol.. 181(16). 5081-5084 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Chang-Kwon Lee: "Identification and in vivo functional analysis of virginiamycin S resistance gene(varS) from Streptomyces virginiae"J. Bacteriol.. 181(10). 3293-3297 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Noriyasu Shikura: "Identification and characterization of 6-dehydro VB-A reductase from Streptomyces antibioticus"FEMS Microbiol. Lett.. 171. 183-189 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Yasuhiro Yamada: "Microbial hormones and microbial chemical ecology" Comprehensive Natural Products Chemistry. 8. 377-413 (1999)

    • Related Report
      1998 Annual Research Report
  • [Publications] Hiroko Nakano: "Gene Replacement Analysis of the Streptomyces virginiae barA Gene Encoding the Butyrolactone Autoregulator Receptor Reveals that BarA Acts as a Repressor in Virginiamycin Biosynthesis" J.Bacteriol.180(13). 3317-3322 (1998)

    • Related Report
      1998 Annual Research Report

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Published: 1998-04-01   Modified: 2016-04-21  

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