Premium
A new method for isolating tyrosine kinase substrates used to identify Fish, an SH3 and PX domain‐containing protein, and Src substrate
Author(s) -
Lock Peter,
Abram Clare L.,
Gibson Toby,
Courtneidge Sara A.
Publication year - 1998
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1093/emboj/17.15.4346
Subject(s) - proto oncogene tyrosine protein kinase src , sh3 domain , biology , sh2 domain , tyrosine kinase , tyrosine phosphorylation , microbiology and biotechnology , tyrosine , phosphorylation , receptor tyrosine kinase , protein tyrosine phosphatase , tyrosine protein kinase csk , actin cytoskeleton , biochemistry , cytoskeleton , signal transduction , cell
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.