DNA-binding activity of adeno-associated virus Rep is required for inverted terminal repeat-dependent complex formation with herpes simplex virus ICP8

J Virol. 2012 Mar;86(5):2859-63. doi: 10.1128/JVI.06364-11. Epub 2011 Dec 28.

Abstract

Herpes simplex virus (HSV) helper functions for (AAV) replication comprise HSV ICP8 and helicase-primase UL5/UL52/UL8. Here we show that N-terminal amino acids of AAV Rep78 that contact the Rep-binding site within the AAV inverted terminal repeat (ITR) are required for ternary-complex formation with infected-cell protein 8 (ICP8) on AAV single-strand DNA (ssDNA) in vitro and for colocalization in nuclear replication domains in vivo. Our data suggest that HSV-dependent AAV replication is initiated by Rep contacting the AAV ITR and by cooperative binding of ICP8 on AAV ssDNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding Sites
  • DNA Helicases / chemistry
  • DNA Helicases / genetics
  • DNA Helicases / metabolism*
  • DNA Replication
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • Dependovirus / chemistry
  • Dependovirus / enzymology*
  • Dependovirus / genetics
  • Dependovirus / physiology
  • HeLa Cells
  • Helper Viruses / genetics
  • Helper Viruses / metabolism
  • Humans
  • Parvoviridae Infections / virology
  • Protein Structure, Tertiary
  • Protein Transport
  • Simplexvirus / genetics
  • Simplexvirus / metabolism*
  • Terminal Repeat Sequences*
  • Viral Proteins / chemistry
  • Viral Proteins / genetics
  • Viral Proteins / metabolism*
  • Virus Replication

Substances

  • DNA-Binding Proteins
  • ICP8 protein, Simplexvirus
  • Viral Proteins
  • DNA Helicases