Mutation causing premature termination of the polyoma virus medium T antigen blocks cell transformation.
We used site-specific mutagenesis to introduce a termination codon, TGA, into the reading frame for the polyoma virus medium T antigen. We induced this mutation in a region of the polyoma genome in which the overlapping coding regions for the large and medium TE antigens are translated in different reading frames. Therefore, the mutation terminated translation of the medium T antigen, but it caused only a single amino acid substitution in the large T antigen and did not affect the small T antigen. Cells infected by the mutant virus produced normal-size small and large T antigens. The infected cells produced a 28,000-dalton fragment of the 48,000-dalton medium T antigen, whose size and tryptic peptide map were consistent with its being a truncated N-terminal fragment terminating at the new termination codon of the mutant. Immunoprecipitates of mutant-infected cell extracts did not show medium-T-antigen-associated protein kinase activity. The mutant virus replicated normally in mouse 3T6 cells and induced cellular DNA synthesis in resting mouse 3T3 cells, but it failed to transform rat or hamster cells, as judged by focus formation and growth in agar. The mutant complemented a tsA mutant which affects the large T antigen for transformation, implying that the mutant defect for transformation was in the medium T antigen. These results imply that the small T antigen and the large T antigen together are insufficient to cause transformation and support the conclusion that the medium T antigen is essential for cell transformation by polyoma virus.
D000818:Animals; D000956:Antigens, Viral / Q000235:genetics*; D000957:Antigens, Viral, Tumor; D002460:Cell Line; D002472:Cell Transformation, Viral; D003062:Codon; D006224:Cricetinae; D008647:Mesocricetus; D051379:Mice; D009154:Mutation; D011120:Polyomavirus / Q000235:genetics*; D014176:Protein Biosynthesis; D051381:Rats
D000956:Antigens, Viral; D000957:Antigens, Viral, Tumor; D003062:Codon