Stable association of activated pp60src with two tyrosine-phosphorylated cellular proteins.
We have identified two phosphotyrosine-containing cellular proteins with relative molecular masses of 130,000 (pp130) and 110,000 (pp110) daltons in chicken embryo cells that coimmunoprecipitated with pp60v-src and activated forms of chicken pp60c-src (pp60(527)F). Most if not all of the tyrosine-phosphorylated forms of pp130 and pp110 could be immunoprecipitated from lysates with any of several src protein-specific monoclonal antibodies directed against at least three spatially distinct epitopes. Consequently, of the more than 15 prominent phosphoproteins detected on immunoblots with phosphotyrosine-specific antibodies, pp130 and pp110 were selectively removed by src protein-specific immunoprecipitation, and their presence in the immunoprecipitates appears to have been due to a direct interaction with activated src proteins. src protein variants that induce different morphological phenotypes were altered in their ability to form detergent-stable complexes with pp130 and pp110 or with pp110 alone. Mutant src proteins, defective for myristylation, showed increased tyrosine phosphorylation of and association with pp110. Expression of src variants with mutations in the A box (pp60dl92/527F) or B box (pp60dl155/527F) of the src homology region induced differences in phosphorylation of pp130 and pp110, as well as changes in their association with variant src proteins. Sequences within the B-box region appeared to be necessary for stable complex formation with pp130 and pp110 and may be involved in the interaction of activated src proteins with cellular substrates.
Reynolds
A B
AB
Department of Microbiology, University of Virginia School of Medicine, Charlottesville 22908.
Kanner
S B
SB
Wang
H C
HC
Parsons
J T
JT
D000818:Animals; D000911:Antibodies, Monoclonal; D001358:Avian Sarcoma Viruses / Q000235:genetics; D002642:Chick Embryo; D004789:Enzyme Activation; D008970:Molecular Weight; D009154:Mutation; D015688:Oncogene Protein pp60(v-src); D010750:Phosphoproteins / Q000276:immunology / Q000378:metabolism*; D010766:Phosphorylation; D019000:Phosphotyrosine; D011494:Protein Kinases / Q000378:metabolism; D012191:Retroviridae Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D014170:Transformation, Genetic; D014443:Tyrosine / Q000031:analogs & derivatives / Q000378:metabolism
D000911:Antibodies, Monoclonal; D010750:Phosphoproteins; D012191:Retroviridae Proteins; D019000:Phosphotyrosine; D014443:Tyrosine; D011494:Protein Kinases; D015688:Oncogene Protein pp60(v-src)