1996 Fiscal Year Final Research Report Summary
The role of phosphoinsitides in cell responses
| Project/Area Number |
07456046
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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 |
応用微生物学・応用生物化学
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| Research Institution | The University of Tokyo |
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Principal Investigator |
FUKUI Yasuhisa University of Tokyo, Faculty of Agriculture and Life Sciences, Professor, 大学院・農学生命科学研究科, 教授 (00181248)
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| Project Period (FY) |
1995 – 1996
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| Keywords | PI-3kinase / phosphorylation / vesicle transport / phosphatidylinositol |
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| Research Abstract |
We have explored the role of Phosphatidylinositol (PI)-3 kinase by two methods. 1) Search for PIP3 (the product of PI-3 kinase) binding protein 2) Expression of activated PI-3 kinase in the cells. For 1), we made a resin that binds PIP3 binding protein specifically. Using this resin, we searched for the PIP3 binding protein. We purified one with the molecular weight of 40kD,cloned the gene for the protein, and analyzed the nucleotide sequence of the gene. We found that the gene encoded a protein with a homology to ARF-GAP protein which is involved in vesicle trafficking suggesting that PI-3 kinase might be involved in membrane trafficking. For 2), we used adenovirus mediated sequence specific recombination. Since constitutively active PI-3 kinase is toxic to the cells, we needed an inducible gene system. In the adenovirus method, genes are completely silent before it is activated by recombination. Often, the gene suppression is leaky when an inducible promotor is used for this type of experiment not allowing us to get a cell line. Using the adenovirus system, we introduced active PI-3 kinase gene into some cell lines. We obteined intersting results suggesting that PI-3 kinase may be involved in cytoskeleton reorganization and membrane trafficking suggesting that we have established cell lines useful of analysis of these kinds of experiments.
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