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Analysis of the Functional Domains of RNA Polymerase

Research Project

Project/Area Number 01480530
Research Category

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

Allocation TypeSingle-year Grants
Research Field 代謝生物化学
Research InstitutionNational Institute of Genetics

Principal Investigator

ISHIHAMA Akira  NAT. INST. GENET., DEPT. MOL. GENET., 分子遺伝研究系, 教授 (80019869)

Co-Investigator(Kenkyū-buntansha) NAGATA Kyosuke  NAT. INST. GENET., DEPT. MOL. GENET., 分子遺伝研究系, 助手 (40180492)
YAMAGISHI Masahiro  NAT. INST. GENET., DEPT. MOL. GENET., 分子遺伝研究系, 助手 (00220252)
藤田 信之  国立遺伝学研究所, 分子遺伝研究系, 助手 (90173434)
Project Period (FY) 1989 – 1990
Project Status Completed (Fiscal Year 1990)
Budget Amount *help
¥6,000,000 (Direct Cost: ¥6,000,000)
Fiscal Year 1990: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1989: ¥3,500,000 (Direct Cost: ¥3,500,000)
KeywordsRNA Polymerase / Functional domain / Transcription factor / Subunit assembly / Transcription regulation / Deletion mutant / Enzyme reconstitution / Escherichia coli / 緊縮制御 / 突然変異 / DNAシ-クエンス
Research Abstract

The DNA-dependent RNA polymerase of Escherichia coli is composed of four major subunits, alpha, beta, beta', and sigma, and carries not only the catalytic activity of RNA synthesis but also complex functions of transmission of genetic information. In order to understand the structural bases for these multiple functions, we carried out a systematic analysis of functional domains on each subunit of E. coli RNA polymerase and obtained the following results.
1) The alpha subunit plays an essential role in the assembly of two large subunits, beta and beta', into enzyme complex. The N-terminal region is required for the sub unit assembly, while the C-terminal domain is needed for transcription activation by position factors which bind DNA upstream of the promoter -35 signal. Mutant RNA polymerases consisting of C-terminal truncated alpha subunits are , however, activated by positive factors which bind to target sites that overlap the promoter -35 region.
2) Four functional sites have been mapped on the beta subunit, including rifampicin bindint site, ppGpp sensitivity control site, promoter selection site and sigma binding site.
3) Putative novel sigma subunits have been identified, using antibodies against synthetic peptides with consensus amino acid sequences common among various sigma subunits. Genes coding for these proteins are being cloned using DNA probes designed based on these amino acid sequences.
4) The functions of these basic subunits are modulated by interaction with accessory transcription factors or modification of enzyme subunits. Sub unit omega is needed for interaction with ppGpp, a regulatory molecule in stringent control.
5) Novel modified forms of RNA polymerase holoenzyme exist in stationary-phase E. coli cells, each recognizing a different set of promoters.
These results altogether indicate that global control of gene transcription takes place mainly by modulating the functions of RNA polymerase.

Report

(3 results)
  • 1990 Annual Research Report   Final Research Report Summary
  • 1989 Annual Research Report
  • Research Products

    (43 results)

All Other

All Publications (43 results)

  • [Publications] Ishihama,A.: "Molecular assembly and functional modulation of Escherichia coli RNA polymerase." Adv.Biophys.26. 19-31 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Igarashi,K.: "Sequence determination of two temperatureーsensitive mutations in the alpha subunit gene (rpoA) of Eschericha coli RNA polymerase." Nucleic Adids Res.18. 5945-5948 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Kobayashi,M.: "Promoter selectivity of Escherichia coli RNA polymerase:Effect of base substitutions in the promoter ー35 region on promoter strength." Nucleic Acids Res.18. 7367-7372 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Igarashi,K.: "Identification of a subunit assembly domain in the alpha subunit of Escherichia coli RNA polymerase." J.Mol.Biol.218. 1-6 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Nakayma,M.: "Identification and characterization of RNA polymerase sigma subunit from Micrococcus luteus." J. Biol. Chem.266. 2911-2916 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Hayward,R.S.: "Functional specialization within the alpha of subunit of Escherichia coli RNA polymerase." J.Mol.Biol.219. (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Ishihama,A.: "Control of Cell Growth and Division" Jpn.Sci.Soc.Press/SpringerーVerlag, 266 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Ishihama,A.: "Structure and Function of Nucleic Acids and Proteins" Raven Press, 336 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Igarashi, K., Fajita, N. and Ishihama, A.: "Promoter selectivity of Escherichia coli RNA polymerase : Omega factor is responsible for the ppGpp sensitivity" Nucleic Acids Res.17. 8755-8765 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Nakayama, M., Fujita, N., Osawa, S. and Ishihama, A.: "Micrococcus luteus, a bacteriu with a high G+C content, contains Escherichia coli-typepromoters." Mol. Gen. Genet.218. 384-389 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Ueshima, R., Fujita, N. and Ishihama, A.: "DNA supercoiling and temperature shift affect the promoter activity of the Escherichia coli rpoH gene encoding the heat-shock sigma subunit of RNA polymerase." Mol. Gen. Genet.215. 185-189 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Igarashi, K., Fujita, N. and Ishihama, A.: "Sequence analysis of two temperature-sensitive mutations in the alpha subunit gene (rpoA) of Escherichia coli RNA polymerase." Nucleic Acids Res.18. 5945-5948 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Igarashi, K., Fujita, N. and Ishihama, A.: "Identificataion of a subunit assembly domain in the alpha subunit of Escherichia coli RNA polymerase." J. Mol. Biol.218. 1-6 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Kobayashi, M., Nagata, K. and Ishihama, A.: "Promoter selectivity of Escherichia coli RNA polymerase : Effect of base substitution in the promoter -35 region on promoter strength." Nucleic Acids Res.18. 7367-7372 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Wada, A., Yamazaki, Y., Fujita, N. and Ishihama, A.: "Structure and probable genetic location of a "ribosome modulation factor" associated with 100S ribosomes in stationary-phase Escherichia coli." Proc. Nat. Acad. Sci. USA. 87. 2657-2661 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Azuma, Y., Yamagishi, M., Ueshima, R. and Ishijima, A.: "Cloning and sequence determination of the Schizosaccharomyces pombe rpb1 gene encoding the largest subunit of RNA polymerase II." Nucleic Acids Res.19. 461-468 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Hayward, R. S., Igarashi, K. and Ishihama, A.: "Functional specialization within the alpha subunit of Escherichia coli RNA polymerase." J. Mol. Biol.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Igarashi, K. and Ishihama, A.: "Bipartite functional organization of the alpha subuni of Escherichia coli RNA polymerase : Involvement of the C-terminal region in the transcription activation by cAMP-CRP." Cell.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Igarashi, K., Hanamura, A., Makino, K., Aiba, Hirofumi, Aiba, Hiroji, Mizuno, T., Nakata, A. and Ishihama, A.: "Bipartite functional organization of the alpha subunit of Escherichia coli RNA polymerase : Two modes of transcription activation positive factors." Cell.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Ozaki, M., Wada, A., Fujita, N. and Ishihama, A.: "Growth phase-dependent conversion of RNA polymerase in Eschelichia coli." Mol. Gen. Genet.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Kajitani, M. and Ishihama, A.: "Identification and sequence determination of the host factor for bacteriophage Q" Nucleic Acids Rec.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Makayama, M., Fujita, N., Osawa, S. and Ishihama, A.: "Identification and characterization of RNA polymerase sigma factor from Microcuccus luteus." J. Biol. Chem. 266. 2911-2916 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Ishihama, A.: "Molecular assembly and functional modulation of Escherichia coli RNA polymerase." Adv. Biophys.Vol. 26. 19-31 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Ishihama, A., Fujita, N., Igarashi, K. and Ueshima, R.: "Structural and functional modulation of Escherichia coli RNA polymerase." Structure and Function of Nucleic Acids and Proteins (Wu, C-W. and Wu, F. Y. -H., eds.145-152 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Ishihama, A.: "Global control of gene expression in bacteria" Control of Cell Growth and Division (Ishihama, A. and Yoshikawa, H., eds.). 121-140 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Ishihama, A.: "Genes for cell growth and division in Eschrichia coli" Control of Cell Growth and Division (Ishihama, A. and Yoshikawa, H., eds.). 199-204 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Ishihama, A., Igarashi, K., Ozaki, M., Hayward, R, S. and Yamagishi, M.: "The promoter selectivity of Escherichia coli RNA polymerase" Gene Expression (Matsubara, K. et al., eds.), Mochida Memorial Found.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Ishihama,A.: "Molecular assembly and functional modulation of <Escherichia>___ー <coli>___ー RNA polymerase." Adv.Biophys.26. 19-31 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Igarashi,K.: "Sequence determination of two temperatureーsensitive mutations in the alpha subunit gene (<rpoA>___ー) of <Eschericha>___ー <coli>___ー RNA polymerase." Nucleic Acids Res.18. 5945-5948 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Kobayashi,M.: "Promoter selectivity of <Escherichia>___ー <coli>___ー RNA polymerase:Effect of base substitutions in the promoter -35 region on promoter strengrh." Nucleic Acids Res.18. 7367-7372 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Igarashi,K.: "Identification of a subunit assembly domain in the alpha subunit of <Escherichia>___ー <coli>___ー RNA polymerase." J.Mol.Biol.218. 1-6 (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] Nakayama,M.: "Identification and characterization of RNA polymerase sigma subunit from <Micrococcus>___ー <luteus>___ー." J.Biol.Chem.266. 2911-2916 (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] Hayward,R.S: "Functional specialization within the alpha subunit of <Escherichia>___ー <coli>___ー RNA polymerase." J.Mol.Biol.219. (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] Ishihama,A.: "Control of Cell Growth and Division" Jpn.Sci.Soc.Press/SpringerーVerlag, 266 (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] Ishihama,A.: "Structure and Eunction of Nucleic Acids and Proteins" Raven Press, 336 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Ishihama,A.: "Promoter selectivity of prokaryotic RNA polymerases." Trends Genet.4. 282-286 (1988)

    • Related Report
      1989 Annual Research Report
  • [Publications] Igarashi,K.: "Promoter selectivity of Escherichi coli RNA polymerase:Omega factor is responsible for the ppGpp sensitivity." Nucleic Acids Res.17. 8755-8765 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Ueshima,R.: "DNA supercoiling and temperature shift affect the promoter activity of the Escherichia coli rpoH gene encoding the heatーshock sigma subunit of RNA polymerase." Mol.Gen.Genet.215. 185-189 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Nakayama,M.: "Micrococcus luteus,a bacterium with a high G+C content,contains Escherichia coliーtype promoters." Mol.Gen.Genet.218. 383-389 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Wada,A.: "Structure and probable genetic map of a ribosome modulation factor associated with 100s ribosome in stationaryーphase Escherichia coli." Proc.Natl.Acad.Sci.USA. (1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] Igarashi,K.: "Functional mapping of the alpha subunit of Escherichia coli RNA polymerase." J.Mol.Biol.(1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] Ishihama,A.: "Switching and Movements of Protein Molecules and Their Assemblies" Jpn.Sci.Soc.Press,Tokyo/Elsevier,Amsterdam, (1990)

    • Related Report
      1989 Annual Research Report
  • [Publications] Ishihama,A.: "Structure and Function of Nucleic Acids and Proteins" Raven Press,New,York, (1990)

    • Related Report
      1989 Annual Research Report

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Published: 1989-04-01   Modified: 2016-04-21  

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