| Project/Area Number |
10215208
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| Research Category |
Grant-in-Aid for Scientific Research on Priority Areas (B)
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| Allocation Type | Single-year Grants |
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| Review Section |
Biological Sciences
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| Research Institution | OKAZAKI NATIONAL RESEACH INSTITUTES (2001)
Okazaki National Research Institutes (1998-2000) |
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Principal Investigator |
YOSHIMORI Tamotsu National Institute of Genetics, Professor, 細胞遺伝研究部門, 教授 (60191649)
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| Co-Investigator(Kenkyū-buntansha) |
HORIUCHI Hisanori Graduate School of Medicine Kyoto University, Assistant Professor, 医学研究科, 助手 (90291426)
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| Project Period (FY) |
1998 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥58,000,000 (Direct Cost: ¥58,000,000)
Fiscal Year 2001: ¥13,500,000 (Direct Cost: ¥13,500,000)
Fiscal Year 2000: ¥16,200,000 (Direct Cost: ¥16,200,000)
Fiscal Year 1999: ¥15,300,000 (Direct Cost: ¥15,300,000)
Fiscal Year 1998: ¥13,000,000 (Direct Cost: ¥13,000,000)
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| Keywords | Autophagy / Autophagosomes / Apg proteins / ES cells / Accumulation of abnormal proteins / Platelet / Granule release / PKCα / エンドソーム / SKD1 / Beclin / Rab4 / 小胞輸送 / 優性阻害変異体 / 輸送再構成形 / 顆粒放出反応 / RabGTPase / AAA型ATPase / GFP / 受容体再循環 / Rab蛋白質 / Aab5 overlay法 / Aab5結合蛋白質 |
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| Research Abstract |
1) Autophagy : We found that binding of the Apg12-Apg5 conjugate to the precursor membranes is required for formation of autophagosomes. Furthermore, we showed that the conjugate forms a large complex (about 700kDa) together with other proteins. We purified one component of the complex and determined its amino acids sequence; it is a 63kDa novel protein, which binds to Apg5 at its N-terminal region. We also demonstrate that GATE 16 and GABARAP, homologues of the autophagosome peripheral membrane protein LC3, are processed in the same way as LC3 and localize to the autophagosomal membranes. Finally, we found that the unfolded protein accumulation is accelerated in the Apg5-deficient cells, suggesting that autophagy is involved in exclusion of abnormal proteins.
2) Secretion in platelet : We established a in vitro reconstitution system for assaying release of serotonin stored in the dense-core granules and von Willebrand factor stored in the α granules by using the platelet whose plasma membrane are permeabilized by Streptolysin-O. Their secretion was dependent on ATP and cytosol. We purified the cytosolic factor required for the secretion and identified it as PKCα. Since PKCα was not sufficient to reconstitute the release, we are trying to identify other factors necessary for the release.
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