Localization of the kinase Ataxia Telangiectasia Mutated to Adenovirus E4 mutant DNA replication centers is important for its inhibitory effect on viral DNA accumulation

Virology. 2019 Jan 15:527:47-56. doi: 10.1016/j.virol.2018.11.003. Epub 2018 Nov 16.

Abstract

Adenovirus (Ad) type 5 (Ad5) E4 deletion mutants including H5dl1007 (E4-) induce a DNA damage response (DDR) that activates the kinase ataxia-telangiectasia mutated (ATM), which can interfere with efficient viral DNA replication. We find that localization of active phosphorylated ATM (pATM) to E4- viral replication centers (VRCs) is important for its inhibitory effect. ATM is necessary for localization of RNF8 and 53BP1 to E4 mutant VRCs, while recruitment of DDR factors Mre11, Mdc1 and γH2AX is ATM-independent, raising the possibility that ATM may affect viral chromatin at VRCs. We assessed E4- and Ad5 chromatin organization by micrococcal nuclease (MN) digestion. A significant fraction of Ad5 DNA is somewhat resistant to MN digestion, whereas E4- DNA is more susceptible. ATM inhibition increases the fraction of E4- DNA that is resistant to MN digestion. Our results address possible mechanisms through which ATM inhibits E4- DNA replication.

Keywords: 53BP1; ATM; Adenovirus; DNA damage response; E4 11 kDa; E4 34 kDa; E4 ORF 3; E4 ORF 6; Micrococcal nuclease; RNF8.

Publication types

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

MeSH terms

  • Adenoviridae / genetics
  • Adenoviridae / metabolism
  • Adenoviridae / physiology*
  • Adenoviridae Infections / metabolism*
  • Adenoviridae Infections / virology
  • Adenovirus E4 Proteins / genetics
  • Adenovirus E4 Proteins / metabolism*
  • Ataxia Telangiectasia Mutated Proteins / antagonists & inhibitors
  • Ataxia Telangiectasia Mutated Proteins / metabolism*
  • Cell Line
  • Chromatin / metabolism
  • DNA Repair
  • DNA, Viral / genetics
  • DNA, Viral / metabolism*
  • DNA-Binding Proteins / metabolism
  • Gene Deletion
  • Humans
  • Micrococcal Nuclease / metabolism
  • Multiprotein Complexes / metabolism
  • Phosphorylation
  • Tumor Suppressor p53-Binding Protein 1 / metabolism
  • Ubiquitin-Protein Ligases / metabolism
  • Virus Replication*

Substances

  • Adenovirus E4 Proteins
  • Chromatin
  • DNA, Viral
  • DNA-Binding Proteins
  • Multiprotein Complexes
  • RNF8 protein, human
  • TP53BP1 protein, human
  • Tumor Suppressor p53-Binding Protein 1
  • Ubiquitin-Protein Ligases
  • Ataxia Telangiectasia Mutated Proteins
  • Micrococcal Nuclease