DNA synthesis and template activity in a mutant of polyoma virus with altered expression of late viral proteins

Abstract

The 3049 strain of polyoma virus overproduces late RNA and proteins. The synthesis and accumulation of virus DNA was measured in cells infected with 3049 and a wild-type virus, lpS, to assess the possible role of gene dosage in this phenomenon. The rate of incorporation of [3H]thymidine into the DNA of 3049 and lpS virus was shown to be identical. The number of genome copies per cell, determined by DNA-DNA reassociation kinetics, was found to be similar in either whole cell or Hirt extracts of cells infected with the two viruses. Viral transcription complexes were isolated by Sarkosyl extraction, and the specific transcriptional activity was measured by incorporation of [3H]UTP. The incorporation of [3H]UTP and the DNA content of transcription complexes were indistinguishable. These results suggest that overproduction of late RNA (and protein) by the 3049 virus is not due to either an increased number of viral genomes per cell or an increased fraction of genomes involved in transcription. These and other data (unpublished) support the conclusion that the altered phenotype is due to a posttranscriptional mechanism.