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Regulatory mechanism of cellular functions of yeast by the ubiquitin-proeasome system.

Research Project

Project/Area Number 10440225
Research Category

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

Allocation TypeSingle-year Grants
Section一般
Research Field 遺伝
Research InstitutionThe University of Tokyo

Principal Investigator

TOH-E Akio  The University of Tokyo, Department of Biological Sciences, Professor, 大学院・理学系研究科, 教授 (90029249)

Co-Investigator(Kenkyū-buntansha) UESONO Yukifumi  The University of Tokyo, Department of Biological Sciences, Assistant Professor, 大学院・理学系研究科, 助手 (30251408)
KIKUCHI Yoshiko  The University of Tokyo, Department of Biological Sciences, Associated, 大学院・理学系研究科, 助教授 (00138124)
Project Period (FY) 1998 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥13,400,000 (Direct Cost: ¥13,400,000)
Fiscal Year 2000: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1999: ¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 1998: ¥8,100,000 (Direct Cost: ¥8,100,000)
Keywords26S proteasome / cell cycle / mitosis / metaphase-anaphase transition / RPN9 / Saccharomyces cerevisiae / two-hybrid screening / プロテアソーム / 蛋白質分解 / Rpn9 / Rpn10 / 蛋白質の集合 / M期中期アレスト / ユビキチン化
Research Abstract

We have isolated the RPN9 gene by two hybrid screening using RPN10 (formerly SUN1) as bait, which encodes a multiubiquitin chain receptor residing in the 26S proteasome. A yeast strain carrying a disrupted allele of RPN9 was found to be temperature sensitive for its growth. At the restrictive temperature, the Δrpn9 strain accumulated multiubiquitinated proteins. When the proteasome fractions separated by the glycerol gradient centrifugation were analysed by native PAGE, we found that the 26S proteasome of the Δrpn9 cells was shifted to lighter fractions than expected and that there was a larger amount of the incomplete proteasome complexes in the fractions. Furthermore, Rpn10 was not detected in the 26S proteasome of Δrpn9 cells. These results indicate that Rpn9 is needed for incorporating Rpn10 into the 26S proteasome and that Rpn9 participates in assembly and/or stability of the 26S proteasome.
Δrpn9 cells were found to be arrested at metaphase ; dumbbell shaped, single nucleus at the isthmuth, unseparated sister chromatids. When a Δrpn9 Δpds1 strain was shifted to a restrictive temperature, the cells stopped growing. No anaphase cells were observed, but sister chrmomatids were separated. Judging from these phenotypes, we anticipate the presence of some proteins which should be degraded by the 26S proteasome during anaphase progression.

Report

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

    (37 results)

All Other

All Publications (37 results)

  • [Publications] Hori.T.,Kato.S.,Sueki.M.,DeMartins.G.N.,Slanghka CA. et al.: "cDNA cloning and functional analysis of p28(Nas6p) and p40.5(Nas7p), two novel regulation subunits of the 26S proteasame"Gene. 216. 113-122 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Takeuchi,J.,Tohe,A.: "Genetic interation between components of the yeast 26S proteasame"Mol.Ger Genet. 262. 145-153 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Takeuchi,J.,Fujimuro.M.,Yokosawa.H.,Tanaka.K.Tohe,A: "Rpn9 is required for efficient assembly of the yeast 26S proteasame"Mol.Cell.Biol.. 19. 6575-6584 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Nishizawa,M.,Kanaya,Y.,Tohe,A.: "Mouse cycling dependent binase(Colk)5 in a functional homologue of a yeast colk, pho85 kinase"J.Biol.Chem.. 274. 33859-33862 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Tone.Y.,Tanahashi.N.,Tanaka.K.,Fujimuro.M.,Yokosawa.H.,Toh-e.A.: "Nob1p, a new essential protein, associates with the 26S proteasome of growing Saccharomyces cerevisiae"Gene. 243. 37-45 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Asakawa.K.,Yosida.S.,Otake.F.,Toh-e.A.: "A novel Functional domain of Cdc15 kinase is required for its interaction with Tem1 GTPase in Saccharomyces cerevisiae"Genetics. (in press). (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Finley, D.et al.: "Unified nomenclature for subunits of the Saccharomyces cerevisiae proteasome regulatory particle."Trend Biochem.Sci.. 23. 244-245 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Hori, T., Kato, S., Saeki, M., DeMartino, G.N., Slaughter, C.A., Takeuchi, J., Toh-e, A.& Tanaka, K.: "cDNA cloning and functional analysis of p28 (Nas6p) and p40.5 (Nas7p), two novel regulatory subunits of the 26S proteasome."Gene. 216. 113-122 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Nishizawa, M., Kawasumi, M., Fujino, M.& Toh-e, A.: "Phosphorylation of Sic1, Cdkinhibitor, by Cdk including Pho85 kinase is required for its prompt degradation."Mol.Biol.Cell. 9. 2393-2405 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Fujimuro, M., Takada, H., Saeki, Y., Toh-e, A., Tanaka, K.& Yokosawa, H.: "Growth-dependent change of the 26S proteasome in budding yeast."Biochem.Biophys.Res.Commun.. 251. 818-823 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Yashiroda, H., Kaida, D., Toh-e, A.& Kikuchi, Y.: "The PY-motif of Bull protein is esential for growth of Saccharmyces cerevisiae under various stress conditions."Gene. 225. 39-46 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Takahashi, Y., Iwase, M., Konishi, M., Tanaka, M., Toh-e, A.& Kikuchi, Y.: "Smt3, a SUMO-1 homolog, is conjugated to Cdc3, a component of septin rings at the mother-budneck in budding yeast."Biochem.Biophys.Res.Commun.. 259. 582-587 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Takeuchi, J.& Toh-e, A.: "Genetic interaction between components of the yeast 26S proteasome : synthetic lethality between a mutation in RPN12 (a lid compnent gene) and thosein RPT1 (an ATPase gene)."Mol.Gen.Genet.. 262. 145-153 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Takeuchi, J., Fujimuro, M., Yokosawa, H., Tanaka, K.& Toh-e, A.: "Rpn9 is required for efficient assenmbly of the yeast 26S proteasome"Mol.Cell.Biol.. 19. 6575-6584 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Tone, Y., Tanahashi, N., Tanaka, K., Fujimura, M., Yokosawa, H.& Toh-e, A.: "Nob1p, a new essential protein, associates with the 26S proteasome of growing Saccharomyces cerevisiae cells."Gene. 243. 37-45 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Saeki, Y., Toh-e, A.& Yokosawa, H.: "Rapid isolation and characterization of the yeast proteasome regulatory complex."Biochem.Biophys.Res.Commun.. 273. 509-515 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Sasaki, T., Toh-e, A., & Kikuchi, Y.: "Yeast Krr1p physically and functionally interacts with a novel essential Kri1p, and both proteins are required for 40S ribosome biogenesis in the nucleolus."Mol.Cell.Biol.. 20. 7971-7979 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Takeuchi, J., & Toh-e, A.: "Genetic dissection of the yeast 26S proteasome : Cell cycle defect caused by the Δrpn9 mutation."Biochemistry. (in press). (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Nishizawa, M., Tanabe, M., Yabuki, N., Kitada, K., & Toh-e, A.: "Pho85 kinase, a yeast cyclin-dependent kinase, regulates the expession of UGP1 encoding UDP-glucose pyrophosphorylase."Yeast. (in press). (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Asakawa, K., Yoshida, S., Ohtake, F., & Toh-e, A.: "A novel functional domain of Cdc15 kinase is required for its interaction with Tem1 GTPase in Saccharomyces cerevisiae."Genetics. (in press). (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Takeuchi,J,Tohe,A: "Genetic dissection of the yeast 26S proteasume"Biochimie. (印刷中). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Toh-e,A.,Asakama,K.,Yoshida,S.: "Spindle checkpoint in budding yeart"J.Microbiol.. (印刷中). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Asakawa,K.,Yoshida,S.,Otake S.,Toh.e.A.: "A novel functional domain of Cdc15 kinase is required for its interaction with Tem1 GTPase in Saccharomyces cerevisiae."Genetics. (印刷中). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Osaka,F.,Saki,M.,Katayama,S.,Toh・e,A et al.: "Covalent modifier NEDD8 is essential for SCF ubiquitious ligase in fission yeast"EMBO J. 19. 3475-3484 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Toh-e,A.,Oguchi,T.: "An improved integration/replacement/disruption method for mutagenics of yeart essential genes"Gene.Genet.Sys.. 75. 33-39 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Toh-e,A and Oguchi,T.: "An improved integration replacement/disruption method"Gene.Genet.Sys.. 75. in press (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] Tone,Y, Tanahashi,N, Tanaka,K, Fujimura,M, Yokosawa, H, and Toh-e,A: "Nob1,a new essenticl protein, associates with the 26S proteasome of growing Saccharomyces cererisiae cells"Gene. in press (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] Toh-e,A. and Oguchi T.: "Las21 participate in extra cellular phenomena in saccharomyces cerevisiae"Gene Genet. Sys.. 74. 241-256 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Nishizawa,M.,Nakaya,Y., and Toh-e,A.: "Mouse cyclin-dependent kinase(Cdk)5 is a functional homologue of a yeast Cdk,Pho85 kinase"J.Biol.Chem.. 274. 33259-33862 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Takeuchi,J.,Fujimuro,M.,Yokosawa,H.,Tanaka,K. and Toh-e A.: "Rpn9 is required for efficient ass*l* by of the yeast 26S protecyme"Mol.Cell Biol.. 19. 6575-6584 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Takeuchi,J.,and Toh-e,A.: "Genetic interaction between components of the yeast 26S protesome synthetic lethality between a mutation in RPN12 and RPT1"Mol.Gene.Genet.. 262. 145-153 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Andoh,T., Kato,T.,Jr., Matsui,Y., Toh-e A: "Phospluslaocifide-specific phospholipase C forms a complex with 14-3-3 proteins and is involved in expression of UV resistance in fission yeast" Mol.Gen.Gevet.258. 139-147 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Fujimuro,M., Tanaka,K., Yokosawa,H., Toh-e,A: "Sonlp is a component of the 26S proteasome of the yeast socchaomy as cerevisiae." FEBS Lett. 423. 149-154 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Fimley,D.et al: "Unified nomeaclature for subunils of the sancheroinyles cerevisice proteasome regulatory particle." Trend.Biochem.Sci.23. 244-245 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Hori,T., Kato,S., Saeki,M., De Marfinoit,N et al: "CDNA Cloning and functional analysis of p28(Nas 6p)and p40.5(Nas 7p), fisc novel segulotory subunits of the 26S proteasome" Gene. 216. 113-122 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Nishizawa,M., Kawasumi,M., Fujino,M., Toh-e,A: "Phosphorylation of Sicl, a Cdk inhibitor, by Cells incladury pho55 kinase is required for its prompt degradation" Mol.Biol.Cell. 9. 2393-2405 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Yashiroda,H., Kaida,D., Toh-e,A., Kikuchi,Y: "The PY-mutif of Bul 1 protein in essential for growth of siccharomyces ceavisiue under various stress conditions" Gene. (in press). (1999)

    • Related Report
      1998 Annual Research Report

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

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