Lamin A/C safeguards replication initiation by orchestrating chromatin accessibility and PCNA recruitment

Lamin A/C, a nuclear lamina protein, is essential for maintaining nuclear architecture, organizing chromatin and preserving genomic stability. However, its role in regulating DNA replication remains unclear. This study investigates how Lamin A/C regulates proper activation of replication origins by orchestrating the chromatin structure and recruitment of proliferating cell nuclear antigen (PCNA). We demonstrate that Lamin A/C stabilizes replication domains (RDs) by restricting chromatin mobility, preserving spatial organization, and maintaining accessibility. Furthermore, Lamin A/C interacts with PCNA and sequesters a pool of PCNA, to regulate its recruitment to replication machinery. The loss of Lamin A/C leads to an increase in chromatin dynamics, RD accessibility, and PCNA availability at RDs, which together trigger excessive activation of replication origins, leading to replication stress and DNA damage. These disruptions prolong the S phase and compromise genome stability, highlighting Lamin A/C as a critical gatekeeper of balanced replication initiation. Our findings reveal Lamin A/C’s dual role in chromatin organization and replication machinery regulation, offering valuable insights into its involvement in replication-associated diseases and potential therapeutic opportunities through targeting replication dynamics.