| Project/Area Number |
05454647
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Cell biology
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| Research Institution | Osaka University |
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Principal Investigator |
NOJIMA Hiroshi Osaka University, RIMD,Department of Molecular Genetics, Professor., 微生物病研究所, 教授 (30156195)
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| Co-Investigator(Kenkyū-buntansha) |
TANAKA Seiji Osaka University, RIMD,Department of Molecular Genetics, Research Associate., 微生物病研究所, 助手 (50263314)
小野 泰子 大阪大学, 微生物病研究所, 教務職員 (70194602)
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| Project Period (FY) |
1993 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥6,600,000 (Direct Cost: ¥6,600,000)
Fiscal Year 1995: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1994: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1993: ¥3,100,000 (Direct Cost: ¥3,100,000)
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| Keywords | CDK5 / Ser / Thr kinase / GST-fusion / BIA core / auixlin / cyclin G / west-western / tensin / 多コピー抑圧遺伝子 / CRE B / Znフィンガー / 機能相補 / 転写制御 / 機能相補クローニング / cDNAライブラリー / cdc2 / cdc13 / cdc10 / 転写因子 / 減数分裂 |
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| Research Abstract |
By west-western hybridization using an anti-cyclin G antibody, we cloned a cDNA encoding a novel association partner of cyclin G.The cDNA encodes a protein harboring a predicted N-terminal Ser/Thr protein kinase catalytic domain with a long C-terminal extension.Hence we named it GAK (cyclin G-associated kinase) . The C-terminal extension shares homology with tensin and auxilin, and contains a leucine zipper. By coimmunoprecipitation and western blotting, we detected and association between GAK and cyclin G in NRK cell extracts, and in extracts of COS cells expressing myc-tagged cyclin G.GAK was also found to coprecipitate with CDK5 and actin, and CDK5 is shown to be associated with cyclin G in NRK extracts. A direct interaction between GAK and cyclin G was detected using immobilized GST-GK and recombinant GST-free cyclin G for BIAcore analysis. Recombinant and immunoprecipitated GAK were found to have kinase activity towards histone H1, but this activity was not enhanced (or decreased) by the presence of cyclin G.In contrast to brain specific expression of auxilin, GAK was found to be ubiquitously expressed. In HeLa cells synchronized from S phase by the thymidine double block, RNA and protein levels of GAK and cyclin G oscillated, peaking at G1 phase, and they showed a similar pattern of oscillation. These results suggest that GAK is somehow involved in cell cycle regulation, forming a complex with cyclin G.
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