Redox regulation of RAD51 Cys319 and homologous recombination by peroxiredoxin 1

Redox Biol. 2022 Oct:56:102443. doi: 10.1016/j.redox.2022.102443. Epub 2022 Aug 24.


RAD51 is a critical recombinase that functions in concert with auxiliary mediator proteins to direct the homologous recombination (HR) DNA repair pathway. We show that Cys319 RAD51 possesses nucleophilic characteristics and is important for irradiation-induced RAD51 foci formation and resistance to inhibitors of poly (ADP-ribose) polymerase (PARP). We have previously identified that cysteine (Cys) oxidation of proteins can be important for activity and modulated via binding to peroxiredoxin 1 (PRDX1). PRDX1 reduces peroxides and coordinates the signaling actions of protein binding partners. Loss of PRDX1 inhibits irradiation-induced RAD51 foci formation and represses HR DNA repair. PRDX1-deficient human breast cancer cells and mouse embryonic fibroblasts display disrupted RAD51 foci formation and decreased HR, resulting in increased DNA damage and sensitization of cells to irradiation. Following irradiation cells deficient in PRDX1 had increased incorporation of the sulfenylation probe DAz-2 in RAD51 Cys319, a functionally-significant, thiol that PRDX1 is critical for maintaining in a reduced state. Molecular dynamics (MD) simulations of dT-DNA bound to a non-oxidized RAD51 protein showed tight binding throughout the simulation, while dT-DNA dissociated from an oxidized Cys319 RAD51 filament. These novel data establish RAD51 Cys319 as a functionally-significant site for the redox regulation of HR and cellular responses to IR.

Keywords: DNA repair; Homologous recombination; Prdx1; RAD51; Sulfenylation.

MeSH terms

  • Adenosine Diphosphate / metabolism
  • Animals
  • Cysteine / metabolism
  • DNA / metabolism
  • DNA Repair
  • Fibroblasts / metabolism
  • Homologous Recombination
  • Humans
  • Mice
  • Oxidation-Reduction
  • Peroxides
  • Peroxiredoxins / genetics
  • Peroxiredoxins / metabolism
  • Poly(ADP-ribose) Polymerase Inhibitors* / pharmacology
  • Poly(ADP-ribose) Polymerases / genetics
  • Rad51 Recombinase* / genetics
  • Rad51 Recombinase* / metabolism
  • Ribose


  • Peroxides
  • Poly(ADP-ribose) Polymerase Inhibitors
  • Adenosine Diphosphate
  • Ribose
  • DNA
  • Peroxiredoxins
  • Poly(ADP-ribose) Polymerases
  • RAD51 protein, human
  • Rad51 Recombinase
  • Rad51 protein, mouse
  • Cysteine