Mechanisms for maintaining genome stability under DNA replication stress

Faithful duplication of the genome is essential for maintaining genome stability, which is vital for cell survival, particularly under DNA replication stress that can lead to cancer. This project combines state-of-the-art single-molecule imaging methods with powerful Xenopus egg extract model system to explore dynamics of DNA replication and repair factors under replicative stress. Our first aim investigates the mechanisms of replication-coupled DNA interstrand crosslink (ICL) processing, critical lesions induced by both endogenous metabolites and exogenous factors such as chemotherapeutics. We will dissect the cellular strategies to process and bypass these lesions. Our second aim focuses on achieving live visualisation of replication fork reversal, a pivotal response to replication stress. This will enable the first real-time observation of replication fork reversal under physiological conditions, shedding light on previously unobservable mechanisms that govern fork stability under stress conditions. Finally, we will explore the interplay between key replication and repair factors that stabilise and restart replication forks under varied stress conditions, thereby enhancing our insight into cellular strategies that safeguard DNA replication fidelity and genome integrity. Together, these aims address critical gaps in our knowledge of the cellular responses to replication stress, with implications for understanding cancer development and improving therapeutic approaches.