The SH2 and SH3 domains of pp60src direct stable association with tyrosine phosphorylated proteins p130 and p110.
Transformation of chicken embryo cells with the tyrosine kinase oncogene src results in the tyrosine phosphorylation of numerous cellular proteins. We have recently generated monoclonal antibodies to individual tyrosine phosphorylated cellular src substrates, several of which are directed to the phosphotyrosine-containing proteins p130 and p110. These proteins form stable complexes with activated variants of pp60src. Mutagenesis of the src homology domains (SH2 and SH3) of activated pp60src resulted in src variants with altered association with p130 and p110. Analysis of these variants showed that the SH3 domain was required for association of p110, while the SH2 domain contained residues necessary for the formation of the ternary complex involving p130, p110 and pp60src. Both the tyrosine phosphorylation status and pp60src association of p130 and p110 appeared to correlate, in part, with the extent of cell transformation. Biochemical analysis demonstrated that p130 and p110 were substrates of both serine/threonine and tyrosine kinases. In addition, p130 was redistributed from the nucleus to cellular membranes upon src transformation, whereas p110, which normally colocalized with cytoskeletal elements, was observed in adhesion plaques (podosomes) in src transformed cells. These data indicate that tyrosine phosphorylation of two different phosphoproteins may play a role during src transformation either by directing their interaction with pp60src, by redirecting subcellular distribution or both.
Kanner
S B
SB
Department of Microbiology, University of Virginia Health Sciences Center, Charlottesville 22908.
Reynolds
A B
AB
Wang
H C
HC
Vines
R R
RR
Parsons
J T
JT
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002642:Chick Embryo; D016296:Mutagenesis; D010449:Peptide Mapping; D010750:Phosphoproteins / Q000737:chemistry*; D010766:Phosphorylation; D019000:Phosphotyrosine; D011487:Protein Conformation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D016392:Proto-Oncogene Proteins pp60(c-src) / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D012689:Sequence Homology, Nucleic Acid; D012694:Serine / Q000378:metabolism; D013329:Structure-Activity Relationship; D013347:Subcellular Fractions / Q000737:chemistry; D013912:Threonine / Q000378:metabolism; D014443:Tyrosine / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism
D000911:Antibodies, Monoclonal; D010750:Phosphoproteins; D019000:Phosphotyrosine; D013912:Threonine; D014443:Tyrosine; D012694:Serine; D011505:Protein-Tyrosine Kinases; D016392:Proto-Oncogene Proteins pp60(c-src)