Herpes simplex virus causes amplification of recombinant plasmids containing simian virus 40 sequences.
Simian virus 40 (SV40) DNA, inserted into a plasmid vector, does not replicate when transfected into baby hamster kidney cells. However, when the recipient cells are superinfected with herpes simplex virus type 1 (HSV-1), extensive amplification of the introduced plasmid occurs. Deletion of the late SV40 region or part of the coding sequences of the small tumour (t) antigen has no effect on the efficiency of amplification, whereas manipulations affecting either the SV40 origin of replication or the integrity of large tumour (T) antigen substantially decrease HSV-induced amplification. Phosphonoacetic acid, an inhibitor of HSV DNA polymerase, strongly inhibits plasmid replication. Also, an HSV-1 mutant with a temperature-sensitive defect in the DNA polymerase gene (tsH) is unable to carry out amplification of test plasmids at the non-permissive temperature. On the other hand, a further mutant (tsS) causes SV40-plasmid amplification independent of the temperature, but this mutant fails to amplify a plasmid with an HSV origin at the non-permissive temperature. Thus, HSV-induced amplification of heterologous DNA is possible in the absence of HSV DNA replication. Since tsS putatively has a defect in the gene coding for an HSV origin-binding protein (UL9), this observation appears plausible. The implications for interaction between herpesviral replication functions and heterologous (possibly cellular) DNA sequences are discussed.
Matz
B
B
Institut für Medizinische Mikrobiologie und Hygiene, Universität Freiburg, F.R.G.
D000818:Animals; D002872:Chromosome Deletion; D004261:DNA Replication / Q000187:drug effects; D004274:DNA, Recombinant; D004279:DNA, Viral / Q000032:analysis / Q000187:drug effects / Q000235:genetics; D005784:Gene Amplification; D005822:Genetic Vectors; D009154:Mutation; D010746:Phosphonoacetic Acid / Q000494:pharmacology; D010957:Plasmids / Q000187:drug effects; D013539:Simian virus 40 / Q000187:drug effects / Q000235:genetics*; D018139:Simplexvirus / Q000187:drug effects / Q000235:genetics*; D014162:Transfection; D014779:Virus Replication / Q000187:drug effects
D004274:DNA, Recombinant; D004279:DNA, Viral; D010746:Phosphonoacetic Acid