Phosphorylation of polyoma middle T antigen and cellular proteins in purified plasma membranes of polyoma virus-infected cells.
We have studied phosphorylation carried out by purified plasma membranes from polyoma virus-infected cells. When isolated membranes are incubated with [gamma-32P]ATP, polyoma virus middle T antigen (mT) becomes phosphorylated on tyrosine. Partial proteolysis mapping shows the same pattern as previously noted for mT labeled in immune complexes. Membranes labeled in vitro were also extracted and immunoprecipitated with anti-T or anti-src antibody. With either antibody, both mT and pp60c-src were brought down and shown to be labeled on tyrosine. The mT of an hr-t mutant (NG59) showed only a trace amount of labeling in membranes under the same conditions. Proteins from infected and uninfected cell membranes labeled in vitro were separated on two-dimensional gels. An acidic 40-kd phosphoprotein was labeled in uninfected cell membranes, but was not seen using membranes from wild-type virus-infected cells. Neither NG59, which encodes a defective but membrane-associated mT, nor a mutant encoding a truncated mT that fails to associate with membranes, alters the level of the 40-kd phosphoprotein in membranes labeled in vitro. These results suggest that mT, acting through pp60c-src and possibly other cellular kinases and phosphatases, can affect cell protein phosphorylation as part of the transformation process.
Ballmer-Hofer
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Benjamin
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TL
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D000952:Antigens, Polyomavirus Transforming; D000957:Antigens, Viral, Tumor / Q000235:genetics*; D002462:Cell Membrane / Q000032:analysis; D002478:Cells, Cultured; D007668:Kidney; D051379:Mice; D008815:Mice, Inbred Strains; D009856:Oncogene Proteins, Viral / Q000235:genetics*; D010766:Phosphorylation; D011120:Polyomavirus / Q000235:genetics* / Q000276:immunology; D011494:Protein Kinases / Q000235:genetics; D011518:Proto-Oncogene Proteins / Q000235:genetics; D016392:Proto-Oncogene Proteins pp60(c-src)
D000952:Antigens, Polyomavirus Transforming; D000957:Antigens, Viral, Tumor; D009856:Oncogene Proteins, Viral; D011518:Proto-Oncogene Proteins; D000255:Adenosine Triphosphate; D011494:Protein Kinases; D016392:Proto-Oncogene Proteins pp60(c-src)