A new method for isolating tyrosine kinase substrates used to identify fish, an SH3 and PX domain-containing protein, and Src substrate.
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.
Lock
P
P
European Molecular Biology Laboratory, Heidelberg, Germany.
Abram
C L
CL
Gibson
T
T
Courtneidge
S A
SA
D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D051379:Mice; D008969:Molecular Sequence Data; D009928:Organ Specificity / Q000235:genetics; D028044:Phosphate-Binding Proteins; D010750:Phosphoproteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis; D013379:Substrate Specificity; D018909:src Homology Domains / Q000235:genetics
C114235:Fish protein, mouse; D028044:Phosphate-Binding Proteins; D010750:Phosphoproteins; D012333:RNA, Messenger; D019000:Phosphotyrosine; D011505:Protein-Tyrosine Kinases