In vitro association and phosphorylation of polyoma virus middle T antigen by cellular tyrosyl kinase activity.
We have observed increased phosphorylation of tyrosine residues on the polyoma virus middle tumor antigen (MTAg) in in vitro kinase assays of the immune complexes immunoprecipitated from lysates of polyoma virus-infected mouse embryo cells to which increasing amounts of uninfected mouse embryo cell lysate had been added. The components from uninfected mouse cells responsible for increased MTAg phosphorylation were localized by subcellular fractionation to the plasma membrane and found to be sensitive to protease digestion, N-ethylmaleimide, and 5'-p-fluorosulfonylbenzoyladenosine inactivation. The majority of the membrane-associated activity responsible for the increased MTAg phosphorylation in these assays could be cleared from lysates of uninfected mouse cell lysates by centrifugation after reaction with Sepharose-bound monoclonal antibodies which recognize pp60c-src. These results suggest that MTAg can associate with cellular tyrosyl kinases in vitro and be phosphorylated by these enzymes in immune-complex kinase assays. The identity of at least one of these cellular tryosyl kinases which can associate with MTAg in vitro is likely to be pp60c-src.
D000241:Adenosine / Q000031:analogs & derivatives / Q000494:pharmacology; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex / Q000032:analysis; D000952:Antigens, Polyomavirus Transforming; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000378:metabolism; D005033:Ethylmaleimide / Q000494:pharmacology; D051379:Mice; D015688:Oncogene Protein pp60(v-src); D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases; D012697:Serine Endopeptidases; D013347:Subcellular Fractions / Q000201:enzymology; D014764:Viral Proteins / Q000276:immunology / Q000378:metabolism*
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D000952:Antigens, Polyomavirus Transforming; D014764:Viral Proteins; C010959:5'-(4-fluorosulfonylbenzoyl)adenosine; D011494:Protein Kinases; D011505:Protein-Tyrosine Kinases; D015688:Oncogene Protein pp60(v-src); D010450:Endopeptidases; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase; D000241:Adenosine; D005033:Ethylmaleimide