Résumé
Replication stress is a major driver of genomic instability and is implicated in the development of diseases such as cancer. It triggers the S-phase checkpoint, a signaling pathway that coordinates the handling of replication obstacles with cell cycle progression. One prominent source of replication stress is the formation of DNA-protein crosslinks on the template, such as those induced by DNA topoisomerase I poisoning by camptothecin (CPT). In this study, we investigated how the S-phase checkpoint responds to CPT-induced replication stress. We show that both activation and timely deactivation of checkpoint signaling are critical for DNA replication completion and cell viability. Using a locus-specific approach, we found that checkpoint signaling is actively dampened at lesion sites. Mechanistically, this attenuation involves the displacement of the checkpoint mediator Rad9 by the DNA repair factors Slx4 and Fun30. This local dampening not only promotes cell cycle progression, but also permits Exo1dependent resection of replication forks stalled by Top1-DNA crosslinks. Controlled resection, in turn, allows homologous recombination factors to access and stabilize the forks, preventing their degradation. In conclusion, we propose that local checkpoint dampening by Slx4 and Fun30 at replication stress sites is a critical mechanism that promotes replication completion and preserves genome stability.