| Project/Area Number |
04044113
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| Research Category |
Grant-in-Aid for international Scientific Research
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| Allocation Type | Single-year Grants |
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| Section | Joint Research |
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| Research Institution | Institute for protein Research, Osaka University |
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Principal Investigator |
NAKAGAWA Hachiro Institute for Protein Research, Osaka University, 蛋白質研究所, 教授 (20029937)
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| Co-Investigator(Kenkyū-buntansha) |
COOPER Jonat Fred Hutchinson Cancer Research Center(U, 室長
PERLMTTER Ro University of Washington(USA), 教授
AIZAWA Shinichi The Institute of Physical and Chemical Research, 副主任 (60073011)
OKUMURA Nobuaki Institute for Protein Research, Osaka University, 蛋白質研究所, 助手 (20224173)
OKADA Masato Institute for Protein Research, Osaka University, 蛋白質研究所, 助手 (10177058)
NAGAI Katsuya Institute for Protein Research, Osaka University, 蛋白質研究所, 助教授 (70029966)
COOPER Jonathan a. Fred Hutchinson Cancer Research Center
PERLMUTTER Roger m. Department of Immunology, University of Washington
ANDRE Veille マッギル大学, 助教授
JONATHAN A C フレッドハッチンソン癌研究所
ROGER M Perl ワシントン大学, 教授
山本 雅 東京大学, 医科学研究所, 教授 (40134621)
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| Project Period (FY) |
1992 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥4,600,000 (Direct Cost: ¥4,600,000)
Fiscal Year 1993: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1992: ¥3,500,000 (Direct Cost: ¥3,500,000)
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| Keywords | Src-family kinases / Protein-tyrosine kinase / Csk / Transgenic mouse / Gene targeting / CSK / src / tyrosine kinasc / phoshorylation |
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| Research Abstract |
Functional analysis of a regulator for Src-family kinases
Src-family kinases are non-receptor type of protein-tyrosine kinases which are localized at the periphery of plasma membrane. It has been suggested that they function as transducers of extracellular signals through physical association with certain type of receptors that lack tyrosine kinase activity, but their genuine functions still remain unknown. The kinase activities of the Src-family kinases are known to be regulated by the phosphorylation of C-terminal regulatory tyrosine residues. In order to shed light on the functions of the Src-family kinases, it seems necessary to figure out the regulation mechanism. Recently we found a novel kinase that can phosphorylate the regulatory tyrosines (termed as Csk) in vitro. A line of evidence obtained in vitro have shown that Csk is the most probable candidate involved in the regulation system, but a question of whether Csk is indeed acting as a regulator for Src-family kinases in vivo
… More
remains to be answered. To address this, we planed to make mutant mice that overexpress Csk in particular tissues or lack Csk, and examine the effects of gain or loss of Csk function on the Src-family kinases.
In University of Washington (Roger Perlmutter lab), we have attempted to generate transgenic mice expressing high level of Csk protein particularly in lymphoid system. However, no mouse produced sufficient amount of Csk protein. We also observed even in cultured cells that it was difficult to gain a cell line overexpressing Csk protein. Thus it was predicted that overexpression of Csk may affect the viability of animals or cells. In Fred Hutchinson Cancer Research Center (Jonathan Cooper Lab), we analyzed the detailed mechanism of the regulation of Src-family kinases mediated by Csk, and found that SH3 domain of Src is also important for the regulation. In Japan (Institute for Protein Research and The Institute of Physical and Chemical Research), we generated Csk-deficient mouse embryos by gene targeting in embryonic stem cells. These embryos were developmentally arrested at the 10 to 12 somite stage and exhibited growth retardation and necrosis in the neural tissues. The kinase activity of Src, Fyn and Lyn in these embryos was greatly enhanced as an apparent consequence of enhanced specific activity. The increase in kinase activity was associated with an increase in tyrosine phosphorylation of several cellular proteins. These findings suggested that Csk indeed act as an indispensable negative regulator of Src-family kinases in vivo. Less
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