Mechanistic insights into RAD51-associated protein 1 (RAD51AP1) action in homologous DNA repair

J Biol Chem. 2012 Apr 6;287(15):12343-7. doi: 10.1074/jbc.C112.352161. Epub 2012 Feb 27.

Abstract

Homologous recombination catalyzed by the RAD51 recombinase is essential for maintaining genome integrity upon the induction of DNA double strand breaks and other DNA lesions. By enhancing the recombinase activity of RAD51, RAD51AP1 (RAD51-associated protein 1) serves a key role in homologous recombination-mediated chromosome damage repair. We show here that RAD51AP1 harbors two distinct DNA binding domains that are both needed for maximal protein activity under physiological conditions. We have finely mapped the two DNA binding domains in RAD51AP1 and generated mutant variants that are impaired in either or both of the DNA binding domains. Examination of these mutants reveals that both domains are indispensable for RAD51AP1 function in cells. These and other results illuminate the mechanistic basis of RAD51AP1 action in homologous DNA repair.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Amino Acid Substitution
  • Conserved Sequence
  • DNA / chemistry
  • DNA Repair*
  • DNA-Binding Proteins / chemistry*
  • DNA-Binding Proteins / genetics
  • HeLa Cells
  • Humans
  • Mutagenesis, Site-Directed
  • Peptide Fragments / chemistry
  • Peptide Fragments / genetics
  • Peptide Mapping
  • Phenotype
  • Protein Binding
  • Protein Structure, Tertiary
  • RNA-Binding Proteins
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Sequence Deletion

Substances

  • DNA-Binding Proteins
  • Peptide Fragments
  • RAD51AP1 protein, human
  • RNA-Binding Proteins
  • Recombinant Proteins
  • DNA