| Project/Area Number |
06454597
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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 |
Biological pharmacy
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| Research Institution | TOKYO UNIVERSITY OF PHARMACY AND LIFE SCIENCE |
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Principal Investigator |
YASUDA Hideyo Tokyo University of Pharmacy and Life Science, School of Life Science, Professor, 生命科学部, 教授 (40111554)
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| Project Period (FY) |
1994 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥7,600,000 (Direct Cost: ¥7,600,000)
Fiscal Year 1996: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1995: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1994: ¥4,000,000 (Direct Cost: ¥4,000,000)
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| Keywords | cdc2 kinase / cdk2 / cyclin / weel kinase / 14-3-3 / cell cycle / myt1キナーゼ / 核移行 / リン酸化 / サイクリン依存性キナーゼ / 転写因子 / PEA3 / weelキナーゼ / 分裂期 |
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| Research Abstract |
To identify proteins which bind to mouse weel kinase, the yeast " two-hybrid"system was used with a mouse cDNA library. Using the carboxyl half of weel kinase, the 14-3-3 zeta protein was isolated. Recombinat 14-3-3 zeta was demonstrated to bind to weel kinase in vitro. When both weel kinase and 14-3-3 zeta were transfected into COS-1 cells, they formed a complex in a cell. The weel kinase phosphorylated by cdc2 kinase also bound to 14-3-3 zeta ptotein. 14-3-3 zeta protein did not affect the activity of weel kinase. Further, cyclin B/cdc2 kinase, weel kinase and 14-3-3 zeta ptotein form a complex. The sequence of weel kinase necessary for the binding was tested by two hybrid system expressing several lengthes of peptides of weel kinase. The entire kinase domain and a sequence in the carboxyl terminus was thought to be necessary for the binding. The function of 14-3-3 zeta protein remained to be elucidated in relationto the regulation of G2 to M phase transition through weel kinase.
The weel kinase phosphorylates only Y-15 residue of cdc2 kinase and T-14 phosphorylation is observed in a intact cell. The other kinase which phosphorylates T-14 of cdc2 kinase must exist. We tried to isolate the T-14 kinase by use of RT-PCR method using DNA sequencest of weel kinase as PCR primers. We isolated mouse mytl kinase which phosphorylate both T-14 and Y-15 in vitro. Cdk2 was also phophorylated by this mytl kinase less than was done by the weel kinase. The weel kinase located in nucleus and mytl kinase located in cytoplasm.
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