Herpes simplex virus type 1 single strand DNA binding protein and helicase/primase complex disable cellular ATR signaling

PLoS Pathog. 2013;9(10):e1003652. doi: 10.1371/journal.ppat.1003652. Epub 2013 Oct 3.

Abstract

Herpes Simplex Virus type 1 (HSV-1) has evolved to disable the cellular DNA damage response kinase, ATR. We have previously shown that HSV-1-infected cells are unable to phosphorylate the ATR substrate Chk1, even under conditions in which replication forks are stalled. Here we report that the HSV-1 single stranded DNA binding protein (ICP8), and the helicase/primase complex (UL8/UL5/UL52) form a nuclear complex in transfected cells that is necessary and sufficient to disable ATR signaling. This complex localizes to sites of DNA damage and colocalizes with ATR/ATRIP and RPA, but under these conditions, the Rad9-Rad1-Hus1 checkpoint clamp (9-1-1) do not. ATR is generally activated by substrates that contain ssDNA adjacent to dsDNA, and previous work from our laboratory has shown that ICP8 and helicase/primase also recognize this substrate. We suggest that these four viral proteins prevent ATR activation by binding to the DNA substrate and obstructing loading of the 9-1-1 checkpoint clamp. Exclusion of 9-1-1 prevents recruitment of TopBP1, the ATR kinase activator, and thus effectively disables ATR signaling. These data provide the first example of viral DNA replication proteins obscuring access to a DNA substrate that would normally trigger a DNA damage response and checkpoint signaling. This unusual mechanism used by HSV suggests that it may be possible to inhibit ATR signaling by preventing recruitment of the 9-1-1 clamp and TopBP1.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Ataxia Telangiectasia Mutated Proteins / genetics
  • Ataxia Telangiectasia Mutated Proteins / metabolism
  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism
  • Cell Cycle Proteins / genetics
  • Cell Cycle Proteins / metabolism
  • Chlorocebus aethiops
  • DNA Helicases / genetics
  • DNA Helicases / metabolism*
  • DNA Primase / genetics
  • DNA Primase / metabolism*
  • DNA, Viral / genetics
  • DNA, Viral / metabolism
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • Exonucleases / genetics
  • Exonucleases / metabolism
  • HeLa Cells
  • Herpesvirus 1, Human / genetics
  • Herpesvirus 1, Human / metabolism*
  • Humans
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism
  • Signal Transduction*
  • Vero Cells
  • Viral Proteins / genetics
  • Viral Proteins / metabolism*

Substances

  • Carrier Proteins
  • Cell Cycle Proteins
  • DNA, Viral
  • DNA-Binding Proteins
  • HUS1B protein, human
  • ICP8 protein, Simplexvirus
  • Nuclear Proteins
  • TOPBP1 protein, human
  • Viral Proteins
  • rad9 protein
  • ATR protein, human
  • Ataxia Telangiectasia Mutated Proteins
  • DNA Primase
  • Exonucleases
  • Rad1 protein, human
  • DNA Helicases