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

EMBO J. 1998 Aug 3;17(15):4346-57. doi: 10.1093/emboj/17.15.4346.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3 Cells
  • Amino Acid Sequence
  • Animals
  • Mice
  • Molecular Sequence Data
  • Organ Specificity / genetics
  • Phosphate-Binding Proteins
  • Phosphoproteins / biosynthesis
  • Phosphoproteins / genetics
  • Phosphoproteins / isolation & purification*
  • Phosphoproteins / metabolism
  • Phosphorylation
  • Phosphotyrosine / metabolism
  • Protein-Tyrosine Kinases / metabolism*
  • RNA, Messenger / biosynthesis
  • Substrate Specificity
  • src Homology Domains* / genetics

Substances

  • Fish protein, mouse
  • Phosphate-Binding Proteins
  • Phosphoproteins
  • RNA, Messenger
  • Phosphotyrosine
  • Protein-Tyrosine Kinases

Associated data

  • GENBANK/AJ007012