title: A mutation in the DNA polymerase accessory factor of herpes simplex virus 1 restores viral DNA replication in the presence of raltegravir.
abstract: Previous reports showed that raltegravir, a recently approved antiviral compound that targets HIV integrase, can inhibit the nuclease function of human cytomegalovirus (HCMV terminase) in vitro. In this study, subtoxic levels of raltegravir were shown to inhibit the replication of four different herpesviruses, herpes simplex virus 1 (HSV-1), HSV-2, HCMV, and mouse cytomegalovirus, by 30- to 700-fold, depending on the dose and the virus tested. Southern blotting and quantitative PCR revealed that raltegravir inhibits DNA replication of HSV-1 rather than cleavage of viral DNA. A raltegravir-resistant HSV-1 mutant was generated by repeated passage in the presence of 200 μM raltegravir. The genomic sequence of the resistant virus, designated clone 7, contained mutations in 16 open reading frames. Of these, the mutations F198S in unique long region 15 (UL15; encoding the large terminase subunit), A374V in UL32 (required for DNA cleavage and packaging), V296I in UL42 (encoding the DNA polymerase accessory factor), and A224S in UL54 (encoding ICP27, an important transcriptional regulator) were introduced independently into the wild-type HSV-1(F) genome, and the recombinant viruses were tested for raltegravir resistance. Viruses bearing both the UL15 and UL32 mutations inserted within the genome of the UL42 mutant were also tested. While the UL15, UL32, and UL54 mutant viruses were fully susceptible to raltegravir, any virus bearing the UL42 mutation was as resistant to raltegravir as clone 7. Overall, these results suggest that raltegravir may be a valuable therapeutic agent against herpesviruses and the antiviral activity targets the DNA polymerase accessory factor rather than the nuclease activity of the terminase.
  
  
  OBJECTIVE
  This paper shows that raltegravir, the antiretrovirus drug targeting integrase, is effective against various herpesviruses. Drug resistance mapped to the herpesvirus DNA polymerase accessory factor, which was an unexpected finding.
authors.affiliation: Department of Microbiology and Immunology, Cornell University, Ithaca, New York, USA College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Zhou
authors.affiliation: Department of Microbiology and Immunology, Cornell University, Ithaca, New York, USA.
authors.forename: Kui
authors.initials: K
authors.lastname: Yang
authors.affiliation: Department of Microbiology and Immunology, Cornell University, Ithaca, New York, USA.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Wills
authors.affiliation: Department of Molecular Biosciences, University of Kansas, Lawrence, Kansas, USA.
authors.forename: Liang
authors.initials: L
authors.lastname: Tang
authors.affiliation: Department of Microbiology and Immunology, Cornell University, Ithaca, New York, USA Jdb11@cornell.edu.
authors.forename: Joel D
authors.initials: JD
authors.lastname: Baines
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line; D002522:Chlorocebus aethiops; D003587:Cytomegalovirus / Q000187:drug effects / Q000235:genetics; D004261:DNA Replication / Q000187:drug effects*; D004279:DNA, Viral / Q000235:genetics; D004259:DNA-Directed DNA Polymerase / Q000235:genetics*; D024882:Drug Resistance, Viral / Q000235:genetics; D005090:Exodeoxyribonucleases / Q000235:genetics*; D015967:Gene Expression Regulation, Viral / Q000187:drug effects; D018259:Herpesvirus 1, Human / Q000187:drug effects* / Q000235:genetics; D018258:Herpesvirus 2, Human / Q000187:drug effects / Q000235:genetics; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D018146:Muromegalovirus / Q000187:drug effects / Q000235:genetics; D009154:Mutation; D011760:Pyrrolidinones / Q000494:pharmacology*; D000068898:Raltegravir Potassium; D014158:Transcription, Genetic / Q000187:drug effects; D014764:Viral Proteins / Q000235:genetics*
chemical_list: D000998:Antiviral Agents; D004279:DNA, Viral; D011760:Pyrrolidinones; D014764:Viral Proteins; D000068898:Raltegravir Potassium; D004259:DNA-Directed DNA Polymerase; D005090:Exodeoxyribonucleases; C043417:DNA polymerase, Simplexvirus