Analysis of complex mutations induced in cells by herpes simplex virus type-1.

The shuttle vector plasmid pZ189 was used as a target for mutagenesis in COS-1 cells. Complex mutations were analyzed in 5 plasmids that were recovered from noninfected cells and 15 plasmids that were recovered from cells that were infected with herpes simplex virus type-1 (HSV-1). Complex mutations in noninfected cells consisted of duplications and rearrangements of plasmid DNA, while those from cells that were infected with HSV-1 included 8 that were enlarged due to insertion of other DNA sequences. Plasmids that contained inserted DNA showed an increase in size of from 118 bp up to around 4500 bp. Maps were constructed based on restriction enzyme digestion, and some or all of each plasmid was examined by DNA sequencing. The inserted DNA was not derived from HSV-1 in any case, since it did not hybridize to DNA from HSV-1 and showed no sequence similarities to the virus. Instead, inserted DNA was found to hybridize to HindIII-digested cellular DNA as a single or double band in 5 plasmids and contained multiple repeat sequences such as alpha satellite, Alu or Kpn repeats in 4 plasmids. In four enlarged plasmids the identity of the inserted sequences could not be determined. The junctions between the shuttle vector and the inserted DNA did not show features of transposable elements and no homology was detected between inserted sequences and the sequence at the insertion site. No preferred site for recombination was detected. Although no similarities were found among the inserted sequences, it is possible that the cellular sequences represent cellular targets for virus-mediated rearrangement. It appears that HSV-1 stimulates nonhomologous recombination between DNA sequences in virus-infected cells.