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Regulation of gene expression by protein kinase in yeast

Research Project

Project/Area Number 04404001
Research Category

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

Allocation TypeSingle-year Grants
Research Field 遺伝学
Research InstitutionUniversity of Tokyo

Principal Investigator

TOH-E Akio  University of Tokyo, Graduate School of Science, Professor, 大学院・理学系研究科, 教授 (90029249)

Co-Investigator(Kenkyū-buntansha) MATSUI Yasushi  Universy of Tokyo, Graduate School of Science, Assistant Professor, 大学院・理学系研究科, 助手 (50229407)
KIKUCHI Yoshiko  University of Tokyo, Graduate School of Science, Associate Professor, 大学院・理学系研究科, 助教授 (00138124)
田中 一馬  東京大学, 理学部, 助手 (60188290)
Project Period (FY) 1992 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥29,000,000 (Direct Cost: ¥29,000,000)
Fiscal Year 1994: ¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1993: ¥6,000,000 (Direct Cost: ¥6,000,000)
Fiscal Year 1992: ¥20,000,000 (Direct Cost: ¥20,000,000)
Keywordsprotein kinase / PHU system / Pho85 / PHO80 / Saccharomyces cerevisiae / プロテインキナーゼ / Saccharomyces cerevisiae / ホスファターゼ / Saccharomyas clrevisial
Research Abstract

PHO85 was first identified as a negative regulator of the PHO system in budding yeast. It encodes a protein kinase that is highly homologous to cyclindependent kinases (CDKs). Since a pho85DELTA mutation causes not only a constitutive expression of the yeast acid phosphatase but defects in normal growth rate and in the utilization of carbon sources, we assume that the Pho85 kinase mediates a signal from the nutrient availability to START progression that is regulated by the Cdc28 kinase. To test this idea, we carried out genetic and biochemical analyzes of the Pho85 kinase.
PHO80 is another negative regulator of the PHO system, functioning together with PHO85 to repress PHO5. By an in vitro kinase assay using kappa-casein as substrate, we found that hyperproduced Pho80 stimulates the Pho85 kinase activity and that an immunoprecipitate containing a tagged Pho80 possessed a kinase activity that was dependent of functional PHO85. These results indicate that the Pho80 protein interacts with the Pho85 kinase to regulate its activity.
To reveal the functional difference between Pho85 and Cdc28, we analyzed the functional domain (s) of the Pho85 kinase. We constructed various point mutants containing mutations in the conserved regions of CDKs. Both a Y18,22F mutation which occurred in the region corresponding to the negative phosphorylation site of cdc2, and a E53A mutation that resides in the PSTAIRE sequence are nonfunctional in vivo as well as in vitro and are dominant negative against the wild type Pho85. On the other hand, YTH (Cdc28 type), FTS,or FAA mutations in F165S166S167, the region corresponding to an active phosphorylation site of cdc2, are functional in vivo. These results indicate that Y18,22 and the PSTAIRE sequence are important for PHO85 function whereas S166 is not, and imply that the function of each domain could be different among CDKs in spite of its conservation.

Report

(4 results)
  • 1994 Annual Research Report   Final Research Report Summary
  • 1993 Annual Research Report
  • 1992 Annual Research Report
  • Research Products

    (30 results)

All Other

All Publications (30 results)

  • [Publications] Fujino,M.,Nishizawa,M.,Yoor,S.-J.,Ozuchi,T.,and Toh.e,A: "Characterization of Pho85 Kinase of Sacharoryces cerevisiae" Phosphate in microorganisms,cellirlar and Molecular Biology Fd.A.Torriani Gorin,F,Yagil,S,Silver. 70-75 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Toh.e,A.: "Pho85" The orutein Kinase facfsbook Ed.Tessa Picknef. in press (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kikuchi,Y.,Oka,Y.,Kobayashi,M.,Uesono,Y.,Toh.e,A.,Kikuchi,A: "A new yeast gene CHTR2,required for growth at high temperature,is needed for recovery from making pheromone-induced GI auert" Mol.Gen.Genet.245. 107-116 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Uesono,Y.,Fujita,A.,Toh.e,A.,Kikuchi,Y.: "The MCSI/SSDI/SRKI/SSLI gene is involved in stable maintenance of chromosune in yeast" Gene. 143. 135-138 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Shirayama,M.,Matsui,Y.,Tanaka,K.,Toh.e,A: "Isulation of a CDC25 Family gene,MSI2/LTE1,as a molticopy suppressor of ijal" Yeast. 10. 451-461 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Shirayama,M.,Matsui,Y.Toh.e,A: "The Yeast TEMI gene,whichencecles a GTP-binding protein,is involved in tenmination of M phase" Mol,Cell,Biol.14. 7476-7482 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Fujino, M., Nishizawa, M., Yoon, S.-J., Oguchi, T., and Toh-e, A.: "Characterization of Pho85 kinase of Saccharomyces cerevisiae." Eds.A.Torriani-Gorini, E.Yagil, and S.Silver ASM,Washington, USA.70-75 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Toh-e, A.: "In The protein kinase factsbook." Ed.Tessa Picknet.Pho85 (in press). (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kikuchi, Y., Oka, Y., Kobayashi, M., Uesono, Y., Toh-e, A., and Kikauchi, A.: "A new yeast gene HTR1, required for growth at high temperature, is needed for recovery from mating pheromone-induced G1 arrest." Mol.Gen.Genet.245. 107-116 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Uesono, Y., Fujita, A., Toh-e, A., and Kikuchi, Y.: "The MCS1/SSD1/SRK1/SSL1 gene is involved in stable maintenance of chromosome in yeast." Gene. 143. 135-138 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Shirayama, M., Matsui, Y., Tanaka, K., and Toh-e, A.: "Isolation of a CDC25yeast family gene, MSI2/LTE1, as a multicopy suppressor of iral." Yeast. 10. 451-461 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Shirayama, M., Matsui, Y., and Toh-e, A.: "The yeast TEM1 gene, which is involved in termination of M phase." Mol.Cell.Biol.14. 7476-7482 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Fujino,M et al.: "Characterization of Pho85 Kinase of Saccharomyces cerevisiae" Phosphate in micro organisms:Cillirlar and Molecular Biology Ed.A.Tottiani.Gorin.E.Yagil.S.Silver. 70-75 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Tohie,A.: "Pho85" The protein Kinase hcctsbook Ed.Tessa Picknet. (in press). (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] Kikuchi,Y.et al.: "A new yeast gene,HTR1,required for growth at high temperature,in needed for recovery from mating pheromone- induced G1 arrest" Mol.Gen.Genet.245. 107-116 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Uesono,Y.et al.: "The MCSI/SSDI/SRKI/SSLI gene is involved in stable maintenance oh chromosure in yeast" Gene. 143. 135-138 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Shirayama,M.et al.: "Isolation of a CDC25 family gene,MSI2/LTE1,as a multicopy suppressor of ital" Yeast. 10. 451-461 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Shirayama,M.et al.: "The Yeast TEMI gene,which encicles a GTP-binding protein,is involved in termination of M phase" Mol.Cell.Biol.14. 7476-7482 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 東江昭夫: "出芽酵母のRas-cAMP経路「酵母から見る真核細胞の最新像」東江ら編" 共立出版, 489 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Yokoo,T.et al.: "Putative phosphoinositicle-specific phospholipase C gene,PLCI,of yeast Saccharomyces cererisiae is important for cell growth." Proc.Natl.Acad.Sci.U.S.A.90. 1804-1808 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Toh-e,A.et al.: "Three yeast genes PIR1,PIR2,and PIR3,containing interaol tandem repeats are related to each other and PIR1 and PIR2 are required for tolerance to heartshock" Yeast. 9. 481-494 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Shirayama,M.et al.: "MSI3,a multicopy suppressor of mutants hyper activated in the RAS-cAMP pataway,encodes on novel HSP70 protein of Saccharomyces cerevisiae." Mol.Gen.Genet.240. 323-332 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Fujimura,K.et al.: "A dominant interfering mutation in RAS1 of Saccharromyces cerevisiae" Mol.Gen.Genet.241. 280-286 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Kominami,K.et al.: "Characterization of the function of the NINI geme product of Saccharomyces cerevisiae." Expt.Cell Res.211(in press). (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] Fujino,M.et al.: "Characterization of PHO85 kinase of Saccharomyces cerevisiae" Pho 93.eds.E.Yagil.,S.Silver. (in press). (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] Uesono,Y., Tokae,M., Tanaka,K., Tohe,A: "Negatae regnlatorsob the PHO syoten of Saahuayas cecevisiae. cbaraeteribation of PHO80 and PHO85genes." Nol, Gen. Genet.231. 426-432 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Okada,H., Tohe,A: "A narel matation occurring in the PHO80 gene snppresseo the PHO40 mntations of Saahawnhces ceieusiae" Curr. Genet.21. 95--99 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Nisogi, H., Kominame,K., sTaraka, K., Tohe,A: "A new essential gene of Sacchauo corevisiae, dofeot of it may result in instability of nucleus." Expt. Cell Res,. 200. 48-57 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Matsae,Y., Tohe,A: "The reart PHOro ahd RHO4 genes, ras superbamily gens, are essential for and grocuth and theu deffect is auppressed by nigh dose of bud-formatin genes CDC42 and BEM1." Mol. Cell. Biol.12. 5690-5699 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Matsni,Y., TOhe,A: "Isolation ayd cbaiac teugation of two novel ias seipen baniy genes in Saccbaroinyes ceuvisial" Gene. 114. 43-49 (1992)

    • Related Report
      1992 Annual Research Report

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Published: 1992-04-01   Modified: 2016-04-21  

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