A viable mouse polyomavirus mutant without immortalizing or transforming activities

Abstract

The polyomavirus mutant, dl1041, has a 375-base pair deletion. It removes most of the sequences that are unique to rodent polyomaviruses and encodes part of the large and middle T-antigens. The mutant was conditionally viable, although both the immortalizing and transforming functions of the T-antigens produced by this mutant were found to be defective. However, the dl1041 mutant was found to be capable of DNA replication in rapidly growing mouse 3T6 cells. In contrast, dl1041 DNA synthesis could not be detected in serum-deprived mouse 3T3 cells. In these cells, the low efficiency of dl1041 DNA replication could be attributed to deficiencies in both large and middle T-antigen, suggesting a link between the mitogenic and oncogenic activities of these proteins. Transfection of growing mouse 3T6 cells with dl1041 DNA resulted in the formation of infectious virus, demonstrating that the dl1041 mutant is able to complete an infection cycle. The ability to activate the viral late promoter in trans was retained by the dl1041 mutant large T-antigen, suggesting that immortalization and trans-activation of the late promoter represent two distinct activities of the protein. An essential element of the immortalizing activity in the large T-antigen polypeptide chain appeared to be in a segment consisting of amino acid residues 136–184, since the dl1041 deletion abolished the activity and the 184 amino acid residue N-terminal dl1354 fragment of large T-antigen retained the activity.