Simian virus 40 T-antigen: identification of tryptic peptides in the C-terminal region and definition of the reading frame.
T-antigen (the simian virus 40 A cistron protein) was purified by immunoprecipitation and electrophoresis on polyacrylamide gels from monkey kidney CV-1 cells infected with simian virus S (SV-S), dl1263, or dl1265 and digested with trypsin. The tryptic peptides, labeled with [35S]methionine, [35S]cysteine, or [3H]proline, were fractionated either by chromatography on Chromobead-P resin or by two-dimensional electrophoresis and chromatography on cellulose thin layers. The T-antigen of SV-S was shown to give rise to a proline-rich (approximately 6 mol of proline) tryptic peptide which was absent in dl1265 T-antigen and hence, on the basis of DNA sequence data, must originate from the C-terminus of the SV-S protein. T-antigen from dl1265, but not SV-S, yielded a cysteine-rich terminal tryptic peptide. The presence of these cysteines caused the protein to be retarded during electrophoresis under the usual conditions in polyacrylamide gels. The T-antigen of dl1263 possessed the proline-rich tryptic peptide; the data are consistent with there being only one peptide altered by the deletion. Both deletion mutants produced a T-antigen that had a higher electrophoretic mobility than SV-S T-antigen but still a larger apparent molecular weight than was predicted by the DNA sequence. The major form of T-antigen found in several lines of 3T3 cells transformed by these mutants was indistinguishable from the T-antigen found in infected cells, and in addition seemed to associate normally with the host-coded 53,000-dalton protein. Except for a minor form of T-antigen with a slightly lower mobility in gels but the same C-terminus, no other polypeptides were detected among the extracted and immunoprecipitated proteins whose electrophoretic mobility was affected by either deletion.
D000956:Antigens, Viral / Q000032:analysis* / Q000235:genetics; D001483:Base Sequence; D002472:Cell Transformation, Viral; D003545:Cysteine / Q000032:analysis; D004279:DNA, Viral / Q000235:genetics*; D009154:Mutation; D010455:Peptides / Q000032:analysis; D011392:Proline / Q000032:analysis; D013539:Simian virus 40 / Q000235:genetics / Q000276:immunology*; D014764:Viral Proteins / Q000032:analysis*
D000956:Antigens, Viral; D004279:DNA, Viral; D010455:Peptides; D014764:Viral Proteins; D011392:Proline; D003545:Cysteine