WRN interacts physically and functionally with the recombination mediator protein RAD52

J Biol Chem. 2003 Sep 19;278(38):36476-86. doi: 10.1074/jbc.M303885200. Epub 2003 May 15.

Abstract

Werner syndrome (WS) is a premature aging disorder that predisposes affected individuals to cancer development. The affected gene, WRN, encodes an RecQ homologue whose precise biological function remains elusive. Altered DNA recombination is a hallmark of WS cells suggesting that WRN plays an important role in these pathways. Here we report a novel physical and functional interaction between WRN and the homologous recombination mediator protein RAD52. Fluorescence resonance energy transfer (FRET) analyses show that WRN and RAD52 form a complex in vivo that co-localizes in foci associated with arrested replication forks. Biochemical studies demonstrate that RAD52 both inhibits and enhances WRN helicase activity in a DNA structure-dependent manner, whereas WRN increases the efficiency of RAD52-mediated strand annealing between non-duplex DNA and homologous sequences contained within a double-stranded plasmid. These results suggest that coordinated WRN and RAD52 activities are involved in replication fork rescue after DNA damage.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Western
  • Cell Line
  • Cell Nucleus / metabolism
  • DNA / chemistry
  • DNA / metabolism
  • DNA Helicases / chemistry*
  • DNA Helicases / physiology*
  • DNA-Binding Proteins / metabolism*
  • Dose-Response Relationship, Drug
  • Exodeoxyribonucleases
  • Fluorescence Resonance Energy Transfer
  • HeLa Cells
  • Humans
  • Immunoblotting
  • Microscopy, Confocal
  • Microscopy, Fluorescence
  • Oligonucleotides / chemistry
  • Plasmids / metabolism
  • Polymerase Chain Reaction
  • Precipitin Tests
  • Protein Binding
  • Rad52 DNA Repair and Recombination Protein
  • RecQ Helicases
  • Recombinant Fusion Proteins / metabolism
  • Recombination, Genetic
  • Substrate Specificity
  • Transfection
  • Two-Hybrid System Techniques
  • Werner Syndrome Helicase

Substances

  • DNA-Binding Proteins
  • Oligonucleotides
  • RAD52 protein, human
  • Rad52 DNA Repair and Recombination Protein
  • Recombinant Fusion Proteins
  • DNA
  • Exodeoxyribonucleases
  • DNA Helicases
  • RecQ Helicases
  • WRN protein, human
  • Werner Syndrome Helicase