A new method for isolating tyrosine kinase substrates used to identify Fish, an SH3 and PX domain-containing protein, and Src substrate

被引:148
作者
Lock, P
Abram, CL
Gibson, T
Courtneidge, SA [1 ]
机构
[1] European Mol Biol Lab, D-69012 Heidelberg, Germany
[2] SUGEN Inc, Redwood City, CA 94063 USA
关键词
cytochalasin D; Fish; PX domain; Src; tyrosine kinase substrate;
D O I
10.1093/emboj/17.15.4346
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We describe a method for identifying tyrosine kinase substrates using anti-phosphotyrosine antibodies to screen tyrosine-phosphorylated cDNA expression libraries. Several potential Src substrates were identified including Fish, which has five SH3 domains and a recently discovered phox homology (PX) domain. Fish is tyrosine-phosphorylated in Src-transformed fibroblasts (suggesting that it is a target of Src in vivo) and in normal cells following treatment with several growth factors. Treatment of cells with cytochalasin D also resulted in rapid tyrosine phosphorylation of Fish, concomitant with activation of Src. These data suggest that Fish is involved in signalling by tyrosine kinases, and imply a specialized role in the actin cytoskeleton.
引用
收藏
页码:4346 / 4357
页数:12
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