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Studies on the structure and function of A-factor receptor based on its X-ray crystallographic structure

Research Project

Project/Area Number 11306008
Research Category

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

Allocation TypeSingle-year Grants
Section一般
Research Field 応用微生物学・応用生物化学
Research InstitutionTHE UNIVERSITY OF TOKYO

Principal Investigator

HORINOUCHI Sueharu  Graduate School of Agriculture and Life Sciences, THE UNIVERSITY OF TOKYO, Professor, 大学院・農学生命科学研究科, 教授 (80143410)

Co-Investigator(Kenkyū-buntansha) OHNISHI Yasuo  Graduate School of Agriculture and Life Sciences, THE UNIVERSITY OF TOKYO, Assistant Professor, 大学院・農学生命科学研究科, 助手 (90292789)
Project Period (FY) 1999 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥30,500,000 (Direct Cost: ¥30,500,000)
Fiscal Year 2000: ¥10,500,000 (Direct Cost: ¥10,500,000)
Fiscal Year 1999: ¥20,000,000 (Direct Cost: ¥20,000,000)
KeywordsA-factor / Streptomyces / A-factor receptor / X-ray chrystallography / microbial hormone / secondary metabolism / morphogenesis
Research Abstract

A chemical signaling molecule A-factor at an extremely low concentration controls secondary metabolism and morphological development in Streptomyces griseus. A rapid method for purification of the A-factor receptor protein was established for crystallization of the receptor. The crystals of the receptor and heavy metal-containing receptor show multiforms, which impeded collection of the difraction data. We are currently collecting data on serenomethione-substituted crystals on Spring 8 at Harima.
More than 20 DNA fragments bound by a transcriptional factor AdpA, one of the targets of the A-factor recepto. Larger DNA fragments covering the isolated fragments were cloned, and their nucleotide sequences were determined. A-factor-dependent transcription of the genes in the cloned fragments was confirmed by S1 nuclease mapping. One of the A-factor-dependent genes was identified as an extracytoplasmic function σ factor of RNA polymerase. This σ factor concerned only with morphological development and not with secondary metabolic function. Additional results are summarized as follows. i) The autophosphorylation of AfsK, a protein serine/threonine kinase, was regulated by means of protein-protein interaction with an AfsK-binding protein KbpA.ii) The whole gene cluster for yellow pigment production, which is under the control of A-factor, was cloed and its nucleotide sequence aws determined. This gene cluster was transcribed by a specific σ^B-like factor. iii) A melanine production genes, rppA and P-450, were cloned. The enzymatic function of RppA was revealed ; it condensated 5 molecules of malonyl-CoA and circulized to form tetrahydroxynaphthalene. Further studied for elucidation of the molecular mechanism of binding and dissociation of A-factor/receptor on the basis of its 3D structure and for revealing each step in the A-factor regulatory cascade will reveal the overall picture of the hormonal control in Streptomyces.

Report

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

    (47 results)

All Other

All Publications (47 results)

  • [Publications] Dae-Kyung Kang: "Possible involvement of cAMP in aerial mycelium formation and secondary metabolism in Streptomyces griseus."Microbiology. 145. 1161-1172 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Yasuo Ohnishi: "The A-factor regulatory cascade leading to streptomycin biosynthesis in Streptomyces griseus : identification of a target gene of the A-factor receptor."Mol.Microbiol.. 34. 102-111 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Takashi Umeyama: "An AfsK/AfsR system involved in the response of aerial mycelium formation to glucose in Streptomyces griseus."Microbiology. 145. 2281-2292 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Tohru Yonekawa: "Involvement of amfC in physiological and morphological development in Streptomyces coelicolor A3(2)."Microbiology. 145. 2273-2280 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Nobutaka Funa: "A new pathway for polyketide synthesis in microorganisms."Nature. 400. 897-899 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Sueharu Horinouchi: "The A-factor regulatory cascade and cAMP in the regulation of physiological and morphological development in Streptomyces griseus"J.Ind.Microbiol.Biotechnol.. 25(in press). (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Sueharu Horinouchi : "Isolation of DNA fragments bound by transcriptional factors, AdpA and ArpA, in the A-factor regulatory cascade"Actinomycetologica. 14. 37-42 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Yasuo Ohnishi : "An oligoribonuclease gene in Streptomyces griseus"J.Bacteriol.. 182. 4647-4653 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Takashi Umeyama : "Genetic and biochemical characterization of a protein phosphatase with dual substrate specificity in Streptomyces coelicolor A3(2)"Gene. 258. 55-62 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Haruka Yamazaki: "An A-factor-dependent extracytoplasmic function sigma factor(σ^<AdsA>)that is essential for morphological development in Streptomyces griseus"J.Bacteriol.. 182. 4596-4605 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Han-Seung Lee: "A σ^B-like factor responsible for carotenoid biosynthesis in Streptomyces griseus"J.Mol.Microbiol.Biotechnol.. 3. 95-101 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Tohru Yonekawa: "A calcium-binding protein with four EF-hand motifs in Streptomyces ambofaciens "Biosci.Biotechnol.Biochem.. 65. 156-160 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 鮒信学: "微生物における新しいポリケタイド生合成経路:新規化合物の生合成工学的手法に利用可能か?"化学と生物. 38. 283-285 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 鮒信学: "微生物のカルコン合成酵素を用いたコンビナトリアル生合成"バイオサイエンスとインダストリー. 58. 568-569 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Sueharu Horinouchi: "Cell-Cell Signaling in Bacteria(分担執筆)"American Society for Microbiology. 490 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] D.-K.Kang, X.-M.Li, K.Ochi and S.Horinouchi.: "Possible involvement of cAMP in aerial mycelium formation and secondary metabolism in Streptomyces griseus."Microbiology. 145. 1161-1172 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Y.Ohnishi, S.Kameyama, H.Onaka and S.Horinouchi.: "The A-factor regulatory cascade leading to streptomycin biosynthesis in Streptomyces griseus : identification of a target gene of the A-factor receptor."Mol. Microbiol.. 34. 102-111 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] N.Funa, Y.Ohnishi, I.Fujii, M.Shibuya, Y.Ebizuka and S.Horinouchi.: "A new pathway for polyketide synthesis in microorganisms."Nature. 400. 897-899 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] T.Umeyama, P.-C.Lee, K.Ueda and S.Horinouchi.: "An AfsK/AfsR system involved in the response of aerial mycelium formation to glucose in Streptomyces griseus."Microbiology. 145. 2281-2292 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] T.Yonekawa, Y.Ohnishi and S.Horinouchi.: "Involvement of amfC in physiological and morphological development in Streptomyces coelicolor A3 (2)."Microbiology. 145. 2273-2280 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] S.Horinouchi, Y.Ohnishi and D.-K.Kang.: "The A-factor regulatory cascade and cAMP in the regulation of physiological and morphological development in Streptomyces griseus."J.Ind. Microbiol. Biotechnol.. 25. 000-000 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] S.Horinouchi, H.Onaka, H.Yamazaki, S.Kameyama and Y.Ohnishi.: "Isolation of DNA fragments bound by transcriptional factors, AdpA and ArpA, in the A-factor regulatory cascade."Actinomycetologica. 14. 37-42 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Y.Ohnishi, Y.Nishiyama, R.Sato, S.Kameyama and S.Horinouchi.: "An oligoribonuclease gene in Streptomyces griseus."J.Bacteriol.. 182. 4647-4653 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] T.Umeyama, A.Naruoka and S.Horinouchi.: "Genetic and biochemical characterization of a protein phosphatase with dual substrate specificity in Streptomyces coelicolor A3 (2)."Gene. 258. 55-62 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] H.Yamazaki, Y.Ohnishi and S.Horinouchi.: "An A-factor-dependent extracytoplasmic function sigma factor (σ^<AdsA>) that is essential for morphological development in Streptomyces griseus."J.Bacteriol.. 182. 4596-4605 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] H.-S.Lee, Y.Ohnishi and S.Horinouchi.: "A σ^B-like factor responsible for carotenoid biosynthesis in Streptomyces griseus."J.Mol. Microbiol. Biotechnol.. 3. 95-101 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] T.Yonekawa, Y.Ohnishi and S.Horinouchi.: "A calcium-binding protein with four EF-hand motifs in Streptomyces ambofaciens."Biosci. Biotechnol. Biochem.. 65. 156-160 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Y.Ohnishi and S.Horinouchi.: "Regulation of secondary metabolism and morphological differentiation by microbial hormones in Streptomyces (in Japanese)."Tanpakusitsu-Kakusan-Kouso. 44. 1552-1561 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] N.Funa and S.Horinouchi.: "A new pathway for polyketide synthesis : its possible application to production of novel compounds (in Japanese)."Kagaku-to-Seibutsu. 38. 283-285 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] N.Funa and S.Horinouchi.: "Combinatorial biosynthesis with a novel chalcone synthase (in Japanese)."Bioscience and Industry. 58. 568-569 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] S.Horinouchi: "γ-Butyrolactones that control secondary metabolism and cell differentiation in Streptomyces."Cell-Cell Signaling in Bacteria, G.M.Dunny and S.C.Winans (eds.), American Society for Microbiology, Washington, D.C.. 193-207 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] S.Horinouchi and Y.Ohnishi: "How A-factor triggers streptomycin biosynthesis in Streptomyces griseus.."Proceedings of International Conference on Asian Network on Microbial Researchers, Chiang Mai, Thailand. 469-477 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Sueharu Horinouchi: "The A-factor regulatory cascade and cAMP in the regulation of physiological and morphological development in Streptomyces griseus"J.Ind.Microbiol.Biotechnol.. 25(in press). (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Sueharu Horinouchi: "Isolation of DNA fragments bound by transcriptional factors, AdpA and ArpA, in the A-factor regulatory cascade"Actinomycetologica. 14. 37-42 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Yasuo Ohnishi: "An oligoribonuclease gene in Streptomyces griseus"J.Bacteriol. 182. 4647-4653 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Takashi Umeyama: "Genetic and biochemical characterization of a protein phosphatase with dual substrate specificity in Streptomyces coelicolor A3(2)"Gene. 258. 55-62 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Haruka Yamazaki: "An A-factor-dependent extracytoplasmic function sigma factor (σ^<AdsA>) that is essential for morphological development in Streptomyces griseus"J.Bacteriol.. 182. 4596-4605 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Han-Seung Lee: "A σ^B-like factor responsible for carotenoid biosynthesis in Streptomyces griseus"J.Mol.Microbiol.Biotechnol.. 3. 95-101 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Tohru Yonekawa: "A calcium-binding protein with four EF-hand motifs in Streptomyces ambofaciens"Biosci.Biotechnol.Biochem.. 65. 156-160 (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] 鮒信学: "微生物における新しいポリケタイド生合成経路:新規化合物の生合成工学的手法に利用可能か?"化学と生物. 38. 283-285 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 鮒信学: "微生物のカルコン合成酵素を用いたコンビナトリアル生合成"バイオサイエンスとインダストリー. 58. 568-569 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Dae-Kyung Kang: "Possible involvement of cAMP in aerial mycelium formation and secondary metabolism in Streptomyces griseus"Microbiology. 145. 1161-1172 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Yasuo Ohnishi: "The A-factor regulatory cascade leading to streptomycin biosynthesis in Streptomyces griseus : identification of a target gene of the A-factor receptor"Mol. Microbiol.. 34. 102-111 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Takashi Umeyama: "An AfsK/AfsR system involved in the response og aerial mycelium formation to glucose in Steptomyces griseus"Microbiology. 145. 2281-2292 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Tohru Yokokawa: "Involvement of amfC in physiological and morphological development in Streptomyces coelicolor A3(2)"Microbiology. 145. 2273-2280 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Sueharu Horinouchi: "The A-factor regulatory cascade and cAMP in the regulation of physiological and morphological development in Strptomyces griseus"J. Ind. Microboil. Biotechnol.. (印刷中). (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] Sueharu Horinouchi: "Cell-Cell Signaling in Bacteria (分担執筆)"American Society for Microbiology. 490(193-207) (1999)

    • Related Report
      1999 Annual Research Report

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Published: 1999-04-01   Modified: 2016-04-21  

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