Fate of viral DNA in nonpermissive cells infected with simian virus 40

Mouse cells are nonpermissive for simian virus 40 (SV40); replication of viral DNA is undetectable and progeny virions are not produced. Infection leads instead to the establishment of stably transformed cell lines in which viral DNA is covalently integrated into cellular DNA. We have followed the fate of SV40 DNA in infected mouse cells to define steps in viral DNA metabolism that precede integration. A novel high molecular weight form of SV40 DNA is synthesized shortly after infection by a process sensitive to the inhibition of DNA replication. This DNA represents polymers in which viral genomes are organized as tandem "head-to-tail" arrays. Recombination can be demonstrated with mutant viruses, but the recombination frequency is not high enough to account for the synthesis of polymers by recombination between infecting genomes. We conclude that polymers are synthesized by DNA replication and that they then recombine with one another. We believe that the polymers also recombine with cellular DNA and are thus the precursor to integrated viral DNA. Such a model accounts directly for the high frequency of tandemly duplicated viral insertions in transformed cells and also leads to experimentally testable predictions.