Ubiquitination of the histone variant mH2A1.2 prevents toxic RAD18 accumulation at a subset of genomic loci upon replication stress. Galloy et al. 2025
Description
Spatiotemporal recruitment of DNA repair factors is crucial to coordinate repair across DNA lesions. MacroH2A1.2 (mH2A1.2), which accumulates in subgenomic regions that are difficult to replicate (e.g., common fragile sites, telomeres, and repeated sequences), selectively promotes homology-driven DNA repair; however, the mechanisms remain elusive. We report an unexpected role for RNF168-mediated mH2A1.2 ubiquitination in preventing aberrant RAD18 recruitment to subgenomic loci experiencing prolonged replication arrest. Biochemical reconstitution revealed that RAD18 cannot bind ubiquitinated mH2A1.2-containing nucleosomes. In cells, loss of mH2A1.2 ubiquitination increases RAD18 and yH2AX levels at collapsed forks and sensitizes cells to replication stress. Depletion of RAD18, fork remodelers, or the endonuclease MUS81 rescues these phenotypes, indicating that mH2A1.2-ubiquitination prevents toxic RAD18 engagement at MUS81-dependent DSBs, which arise at collapsed forks in difficult to replicate sites experiencing prolonged arrest. Our findings highlight the detrimental consequences of inappropriate DNA processing by MUS81 at these loci. Raw and unprocessed images of all immunoblots, SDS-PAGE, native gels and immunofluorescence images are provided. High-resolution images or any additional information required to reanalyze the data reported in this paper, including large images of immunofluorescence used for quantification, are available from the lead contact upon request.
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Institutions
- CHU de Quebec-Universite Laval
- Universite Laval