EXAMINATION OF REGULATROY MECHANISMS FOR CELL FUNCTIONS BY THE GOLGI APPARATUS
| Project/Area Number |
15570156
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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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 | KANAZAWA UNIVERSITY |
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Principal Investigator |
NAKAMURA Nobuhiro KANAZAWA UNIVERSITY, GRADUATE SCHOOL OF NATURAL SCIENCE AND TECHNOLOGY, ASSOCIATE PROFESSOR, 自然科学研究科, 助教授 (50294955)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2004: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 2003: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Keywords | GOLGI APPARATUS / PHOSPHORYLATION / KINASE / GROWTH SIGNAL |
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| Research Abstract |
We have shown that a 277th amino acid residue of GRASP65 (S277) is phosphorylated in interphase cells and the phosphorylation signal is markedly enhanced by the growth factor treatment including EGF. ERK is activated by the EGF induced growth factor signal and the activated ERK phosphorylates S277 directly. We further found that S277 is heavily phosphorylated during M phase and analyzed the molecular mechanism for this up regulation of the phosphorylation. The amino acid sequence around S277 (PGSPG) is well conserved among mammalian CTRASP65 homologues. This sequence is well fitted with the target sequence of cdk1/cyclinB. S277 was strongly phosphorylated by a cytoplasmic extract of M phase cells and this was completely inhibited by roscovitine, a cdk specific inhibitor. S277 was also phosphorylated by an ERK inactive cytoplasmic extract of M phase cells that was prepared in the presence of U0126, a MEK inhibitor. These results strongly suggested that cdk1/cyclinB, and not ERK, is responsible for the phosphorylation of S277 in M phase. Surprisingly, the mitotic entry was strongly inhibited by the microinjection of purified GRASP65 without N-terminal myristoylation (Δm-GRASP65). This was not observed by the microinjection of Δm-GRASP65 in which S277 was changed with alanine. These results suggested that Δm-GRASP65 interact with some cytoplasmic factors and inhibits the mitotic entry. We have found that Plk1 specifically binds to phosphorylated S277 region of GRASP65 and there are some cytoplasmic factors that bind to unphosphorylated S277 region of GRASP65.
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Report
(3 results)
Research Products
(11 results)
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[Journal Article] Convergence of cell cycle regulation and growth factor signals on GRASP65.2005
Author(s)
Yoshimura, S., Yoshioka, K., Barr, F.A., Lowe, M., Nakayama, K., Ohkuma, S., Nakamura, N.
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Journal Title
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Dynamics of Golgi matrix proteins after a block of ER to Golgi transport.2004
Author(s)
Yoshimura, S., Yamamoto, A., Misumi, Y., Sohda, M., Barr, F.A., Fujii, G., Shakoori, A., Ohno, H., Mihara, K., Nakamura, N.
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Journal Title
J.Biochem. 135
Pages: 201-216
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Identification of a five-pass transmembrane protein family localizing in the Golgi apparatus and the ER.2003
Author(s)
Shakoori, A., Fujli, G., Yoshimura, S., Kitamura, M., Nakayama, K., Ito, T., Ohno, H., Nakamura, N.
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Journal Title
Biochem.Biophys.Res.Commun. 312
Pages: 850-857
Description
「研究成果報告書概要(欧文)」より
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