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Redox regulation of RAD51 Cys319 and homologous recombination by peroxiredoxin 1
Journal article   Open access   Peer reviewed

Redox regulation of RAD51 Cys319 and homologous recombination by peroxiredoxin 1

John J. Skoko, Juxiang Cao, David Gaboriau, Myriam Attar, Alparslan Asan, Lisa Hong, Candice E. Paulsen, Hongqiang Ma, Yang Liu, Hanzhi Wu, …
Redox biology, Vol.56, p.102443
10/2022
DOI: 10.1016/j.redox.2022.102443
PMCID: PMC9450138
PMID: 36058112
url
https://doi.org/10.1016/j.redox.2022.102443View
Published (Version of record) Open Access

Abstract

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. [Display omitted] •A functionally-significant Cys319 was identified in RAD51 that possesses nucleophilic characteristics.•RAD51 Cys319 plays a central role in RAD51-mediated repair of DNA double strand breaks (DSB).•Loss of peroxiredoxin 1 (PRDX1) impairs DNA DSB repair by homologous recombination and results in DNA damage.•PRDX1 is critical for maintaining RAD51 Cys319 in a reduced state.•Molecular dynamic (MD) simulations suggest ssDNA to dissociate from sulfenylated and not reduced RAD51 Cys319.
DNA Repair Homologous recombination Prdx1 RAD51 Sulfenylation

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