| Project/Area Number |
14380336
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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 | OKAYAMA UNIVERSITY |
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Principal Investigator |
TAKEI Kohji Okayama University, Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Professor, 大学院・医歯薬学総合研究科, 教授 (40322226)
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| Co-Investigator(Kenkyū-buntansha) |
YAMADA Hiroshi Okayama University, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Lecturer, 大学院・医歯薬学総合研究科, 講師 (80325092)
絹田 正裕 岡山大学, 大学院・医歯学総合研究科, 講師 (40135942)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥11,100,000 (Direct Cost: ¥11,100,000)
Fiscal Year 2004: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2003: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2002: ¥6,000,000 (Direct Cost: ¥6,000,000)
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| Keywords | endocytosis / vesicular traffic / cell-free system / live imaging / lyposomes / dynamin / amphiphysin / 神経 / 脳 / イノシトールリン脂質 / PIP_2 |
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| Research Abstract |
Regarding to regulatory mechanisms of endocytosis, the followings were revealed.
1.Regulation of endocytosis by Amphiphysin 1
Dynamin-dependent endocytosis can be reconstituted in vitro by incubating large unilammelar liposomes with brain cytosol or dynamin in presence of GTP. Using amphipshyin knockout brain cytosol in this experimental system, it was clarified that amphiphysin 1 stimulates dynamin GTPase activity and thereby enhances dynamin-dependent vesicle formation. This effect required both BAR domain and SH3 domain of amphiphysin 1. Low membrane curvature of large liposomes was also requisite for the stimulatory effect of amphiphysin 1.
2.Regulation of endocytosis cdk5-dependent phosphorylation
Both dynamin 1 and amphiphysin 1 are phosphorylated by cyclin dependent kinase 5 (cdk5). Incubation of liposomes with phosphorylated dynain and phosphorylated amphiphysin 1 in presence of GTP resulted in few vesicle formation, whereas dephophorylated proteins massively generated vesicles. Thus, endocytosis is likely to be regulated by cdk5-dependent phosphorylation.
3.Localization of dynamin 2 and dynamin3
Distinct localization of dynamin 2 and dynamin3 in Sertoli cells was revealed suggesting different functions of these isoforms.
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