Sessa, GaetanaGómez-González, BelénSilva, SoniaPérez-Calero, CarmenBeaurepere, RomaneBarroso, SoniaMartineau, SylvainMartin, CharlotteEhlén, ÅsaMartínez, Juan SLombard, BérangèreLoew, DamarysVagner, StephanAguilera, AndrésCarreira, Aura2023-02-092023-02-092021-02-26http://hdl.handle.net/10668/17243The BRCA2 tumor suppressor is a DNA double-strand break (DSB) repair factor essential for maintaining genome integrity. BRCA2-deficient cells spontaneously accumulate DNA-RNA hybrids, a known source of genome instability. However, the specific role of BRCA2 on these structures remains poorly understood. Here we identified the DEAD-box RNA helicase DDX5 as a BRCA2-interacting protein. DDX5 associates with DNA-RNA hybrids that form in the vicinity of DSBs, and this association is enhanced by BRCA2. Notably, BRCA2 stimulates the DNA-RNA hybrid-unwinding activity of DDX5 helicase. An impaired BRCA2-DDX5 interaction, as observed in cells expressing the breast cancer variant BRCA2-T207A, reduces the association of DDX5 with DNA-RNA hybrids, decreases the number of RPA foci, and alters the kinetics of appearance of RAD51 foci upon irradiation. Our findings are consistent with DNA-RNA hybrids constituting an impediment for the repair of DSBs by homologous recombination and reveal BRCA2 and DDX5 as active players in their removal.enAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/BRCA2DNA double-strand breaksDNA-RNA hybridsR-loopshomologous recombinationBRCA2 ProteinCell Line, TumorDEAD-box RNA HelicasesDNA Breaks, Double-StrandedHEK293 CellsHumansNucleic Acid HeteroduplexesProtein BindingRecombinational DNA Repairresearch article33634895open access10.15252/embj.20201060181460-2075PMC8013831https://idus.us.es/bitstream/11441/110909/1/2021.EMBO%20J-Sessa.pdfhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC8013831/pdf