Tyrosine phosphorylation of a 120-kilodalton pp60src substrate upon epidermal growth factor and platelet-derived growth factor receptor stimulation and in polyomavirus middle-T-antigen-transformed cells.

Kanner SB, Reynolds AB, Parsons JT
Mol Cell Biol. 1991 11 (2): 713-20

PMID: 1703631 · PMCID: PMC359722 · DOI:10.1128/mcb.11.2.713

The monoclonal antibody 2B12 is directed toward p120, a 120-kDa cellular protein originally identified as a protein tyrosine kinase substrate in cells expressing membrane-associated oncogenic variants of pp60src. In this report, we show that p120 was tyrosine phosphorylated in avian cells expressing membrane-associated, enzymatically activated variants of c-src, including variants having structural alterations in the src homology regions 2 and 3. In contrast, p120 was not tyrosine phosphorylated in cells expressing enzymatically activated, nonmyristylated pp60src. Furthermore, p120 was tyrosine phosphorylated in avian cells expressing middle T antigen, the transforming protein of polyomavirus, as well as in rodent cells stimulated with either epidermal growth factor (EGF) or platelet-derived growth factor. Analysis of the time course of p120 tyrosine phosphorylation in EGF-stimulated cells revealed a rapid onset of tyrosine phosphorylation. In addition, both the extent and duration of p120 phosphorylation increased when cells overexpressing the EGF receptor were stimulated with EGF. Biochemical analysis showed that p120 (in both normal and src-transformed cells) was membrane associated, was myristylated, and was phosphorylated on serine and threonine residues. Hence, p120 appears to be a substrate of both nonreceptor- and ligand-activated transmembrane receptor tyrosine kinases and of serine/threonine kinases and is perhaps a component of both mitogen-stimulated and tyrosine kinase oncogene-induced signaling pathways.

MeSH Terms (21)

Animals Antigens, Polyomavirus Transforming Cell Line Cell Transformation, Neoplastic Chick Embryo ErbB Receptors Humans Kinetics Molecular Weight Moloney murine leukemia virus Myristic Acid Myristic Acids Palmitic Acid Palmitic Acids Phosphates Phosphorylation Platelet-Derived Growth Factor Proto-Oncogene Proteins pp60(c-src) Receptors, Cell Surface Receptors, Platelet-Derived Growth Factor Tyrosine

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