1996 Fiscal Year Final Research Report Summary
Functional analyzes of molecular chaperons concerning on protein translocation across the ER membrane
| Project/Area Number |
07458254
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Cell biology
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| Research Institution | Nara Institute of Science and Technology |
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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)
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| 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)
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| Project Period (FY) |
1995 – 1996
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| Keywords | HSP70 / Stress Response / BiP / yeast / HSE / Microtubule / Tubulin |
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| 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.
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[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)
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