Inhibition of the herpes simplex virus type 1 DNA polymerase induces hyperphosphorylation of replication protein A and its accumulation at S-phase-specific sites of DNA damage during infection

J Virol. 2005 Jun;79(11):7162-71. doi: 10.1128/JVI.79.11.7162-7171.2005.

Abstract

The treatment of mammalian cells with genotoxic substances can trigger DNA damage responses that include the hyperphosphorylation of replication protein A (RPA), a protein that plays key roles in the recognition, signaling, and repair of damaged DNA. We have previously reported that in the presence of a viral polymerase inhibitor, herpes simplex virus type 1 (HSV-1) infection induces the hyperphosphorylation of RPA (D. E. Wilkinson and S. K. Weller, J. Virol. 78:4783-4796, 2004). We initiated the present study to further characterize this genotoxic response to HSV-1 infection. Here we report that infection in the presence of polymerase inhibitors triggers an S-phase-specific response to DNA damage, as demonstrated by induction of the hyperphosphorylation of RPA and its accumulation within viral foci specific to the S phase of the cell cycle. This DNA damage response occurred in the presence of viral polymerase inhibitors and required the HSV-1 polymerase holoenzyme as well as the viral single-stranded-DNA binding protein. Treatment with an inhibitor of the viral helicase-primase did not induce the hyperphosphorylation of RPA or its accumulation in infected cells. Taken together, these results suggest that the S-phase-specific DNA damage response to infection is dependent on the specific inhibition of the polymerase. Finally, RPA hyperphosphorylation was not induced during productive infection, indicating that active viral replication does not trigger this potentially detrimental stress response.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Chlorocebus aethiops
  • DNA Damage*
  • DNA, Viral / biosynthesis
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / metabolism*
  • DNA-Directed DNA Polymerase / physiology
  • Enzyme Inhibitors / pharmacology
  • Exodeoxyribonucleases / antagonists & inhibitors*
  • Exodeoxyribonucleases / physiology
  • Herpes Simplex / metabolism
  • Herpes Simplex / pathology
  • Herpes Simplex / virology
  • Herpesvirus 1, Human / drug effects
  • Herpesvirus 1, Human / enzymology*
  • Herpesvirus 1, Human / pathogenicity
  • Herpesvirus 1, Human / physiology
  • Nucleic Acid Synthesis Inhibitors* / pharmacology
  • Phosphorylation
  • Replication Protein A
  • S Phase
  • Vero Cells
  • Viral Proteins / antagonists & inhibitors*
  • Viral Proteins / physiology
  • Virus Replication

Substances

  • DNA, Viral
  • DNA-Binding Proteins
  • Enzyme Inhibitors
  • Nucleic Acid Synthesis Inhibitors
  • Replication Protein A
  • Viral Proteins
  • DNA-Directed DNA Polymerase
  • Exodeoxyribonucleases
  • DNA polymerase, Simplexvirus