Proteome dynamics at stalled replication forks reveal a coordinated recruitment of repair factors

ABSTRACT: The faithful replication of DNA is a fundamental cellular process that ensures the transfer of genetic information during cell division. Genome duplication takes place during S phase and requires the dynamic and coordinated recruitment of multiple proteins at the replication forks. The instability and collapse of the replication fork can occur when cells are exposed to different genotoxic stresses, which can be overcome by the action of DNA repair pathways. Studying the proteomic interactions implicated in those events is thus a crucial step in understanding the complex biological mechanisms involved in DNA replication and repair. Proximity-dependent biotinylation coupled with mass spectrometry identification of proteins (BioID2) was used on 17 proteins from four different complexes known to play a role during DNA replication and repair to study the assembly and recruitment of proteins at stalled replication forks caused by treatment with hydroxyurea. This analysis revealed a vast interaction network of 108 proteins modulated in the presence of hydroxyurea; 45 being enriched and 63 depleted. Interestingly, 11 of them were found to be both enriched and depleted depending on the complex considered, suggesting a dynamic reorganization of the players and their interactions (?). Furthermore, the analysis identified several poorly characterized proteins, thus uncovering new players in the cellular response to DNA replication arrest. Overall, this collection of replication fork proteomes enables large-scale identification of the mechanisms of assembly and disassembly of protein complexes at the sites of stalled replication forks and provides a new framework to understand how cells respond to obstacles on DNA template, causing fork stalling or collapse.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell, Cell Culture

DISEASE(S): Colon Cancer,Breast Cancer

SUBMITTER: Dominique Levesque  

LAB HEAD: François-Michel Boisvert

PROVIDER: PXD037136 | Pride | 2026-02-04

REPOSITORIES: Pride