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Functional analyzes of molecular chaperons concerning on protein translocation across the ER membrane

Research Project

Project/Area Number 07458254
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Cell biology
Research InstitutionNara Institute of Science and Technology

Principal Investigator

KOHNO Kenji  Nara Institute of Science and Technology Research and Education Center for Genetic Information, Professor, 遺伝子教育研究センター, 教授 (50142005)

Co-Investigator(Kenkyū-buntansha) ISURU Akio  Nara Institute of Science and Technology Research and Education Center for Genet, 遺伝子教育研究センター, 助手 (80273861)
KIMATA Yukio  Nara Institute of Science and Technology Research and Education Center for Genet, 遺伝子教育研究センター, 助手 (60263448)
Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1996: ¥2,100,000 (Direct Cost: ¥2,100,000)
KeywordsHSP70 / Stress Response / BiP / yeast / HSE / Microtubule / Tubulin / hsp70 / Ssalp / Green Fluorescent Protein / 核の配分
Research Abstract

Bip is the member of HSP70 family that is localized in the lumen of the endoplasmic reticulum (ER) and plays an important role in translocation of nascent proteins across the ER membrane and their subsequent folding and assembly in the ER lumen. BiP is induced at the transcriptional level by various environmental stresses such as high temperatue, block of the protein glycosylation, intracellular Ca^<2+> disturbance, the addition of the recuded agents, those are leading to the accumulation of unfolded proteins in the ER.We found that depletion of intracellular Ssa1p induced KAR2 gene expression at the transcriptional level. By analyzing internal deletion mutants of the KAR2 promoter, heat shock element (HSE) is necessary for KAR2 gene induction in response to the depletion of Ssa1p. a temperature-sensitive ssal mutant transformed with HSE-CYCl-lacZ fusion vector exhibited the highly induced beta-galactosidase activity compared with that of wild-type control cells at restrictive temperature. These results show that the consentration of functional Ssalp in the cytosol participates in the regulation of KAR2 gene expression through HSE-mediated pathway and also support the idea that SSAl gene expression is auto-regulated. Another findings is that, after shift to the restrictive temperature (37゚C), ssal temperature-sensitive mutant cells showed abnormal distribution of nucleus and accumulated as large-budded cells with 2N DNA contents accompanied by aberrant microtubule structure. This result suggests llthat Ssalp is involved in function of microtubules.

Report

(3 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] M. Oka: "Saccharomyces cerevisiae KAR2 (BiP) gene expression is induced by loss of cytosclic HSP70/Ssalp through heat shock element-medirted pathnay" J. Biochemistry. 121. 578-584 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Y. Kimata: "A novel mutation which enhances the fluorescence of green flucrescent protein at high tomperatures" Biochem. Biophys. Res. Commun.232. 69-73 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] H. Hayes: "Localization of ZNF164, ZNF146, GGTA1, SOX2, PRLR and EEF2 on homoeologs cattle, sheep, and goat chemosomes by flurescent in situ hgbriclizatio and comprison" Cyto genet. Cell Genot.72. 342-346 (1996)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] T. Shimamoto: "Expression and functional analyses of Dxpa gene the Drasophila homolog of human excision repair gene XPA" J. Biol. Chem.270. 22452-22459 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] C. R. Lim: "Thermosonsitivity of green fhiorescent protein fluorescence utilized to rereal novel nuclear-like compartments in a mutant mucletpon'n NSP1" J. Biochem.118. 13-17 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] M.Oka, Y.Kimata, K.Mori and K.Kohno: "Saccharomyces cerevisiae KAR2 (BiP) gene expression is induced by loss of ctyosolic HSP70/Ssalp through a heat shock element-mediated pathway" J.Biochem. 121. 578-584 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Y.Kimata, M.Iwaki, C.R.Lim and K.Kohno: "A novel mutation which enhances the fluorescence of green flourescent protein at high temperatures" Biochem.Biophys.Res.Commun.232. 69-73 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] H.Hays, C.Le Chalony, G.Goubin, D.Mercier, E.Payn, C.Bignon, and K.Kohno: "Localization of ZNF164, ZNF146, GGTA1, SOX2, PRLR and EEF2 on homoeologous cattle, sheep and goat chromosomes by fluorescent in situ hybridization and comparison with the human gene map" Cytogenet.Cell Genet.72. 342-346 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] T.Shimamoto, T.Tanimura, Y.Yoneda, Y.Kobayakawa, K.Sugasawa, F.Hanaoka, M.Oka, Y.Okada, K.Tanaka, and K.Kohno: "Expression and functional analyzes of Dxpa gene the Drosophlia homolog of human excision repair gene XPA" J.Biol.Chem.270. 22452-22459 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] C.R.Lim, Y.Kimata, M.Oka, K.Nomaguchi, and K.Kohno: "Thermosensitivity of green fluorescent protein fluorescence utilized to reveal novel nuclear-like compartments in a mutant nucleoporin NSP1" J.Biochem.118. 13-17 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] M.Oka: "Sacchromyces cerevisiae KAR2(BiP)gene expression is induced by loss of cytosolic HSP70%Ssalp though a heat shock element-mediated pathway" J.Biochemistry. 121. 578-584 (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Y.Kimata: "A novel mutation which enhances the fluorescence of green fluorescent protein at high temperature." Biochem.Biophys.Res.Commun.(in press). (1997)

    • Related Report
      1996 Annual Research Report
  • [Publications] Lim,C.R.,et al.: "The rmosensitivity of green fluorescent protein fluorescence utilized to reveal novel nuclear-like compnrtments in a mutant nuclcopcrin NSP1." J.Biochem.118. 13-17 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Shimamoto,T.,et al.: "Expression and functional analyses of Dxpa gene ,the Drosophila homolog of human excision repaiy gene XPA." J.Biol Chem.270. 22452-22459 (1995)

    • Related Report
      1995 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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