Rebecca S Rivard, Ya-Chu Chang, Ryan L Ragland, Yee-Mon Thu, Muzaffer Kassab, Rahul Shubhra Mandal, Susan K Van Riper, Katarzyna Kulej, LeeAnn Higgins, Todd M Markowski, David Shang, Jack Hedberg, Luke Erber, Benjamin Garcia, Yue Chen, Anja-Katrin Bielinsky, Eric J Brown
Innovative methods to retrieve proteins associated with actively replicating DNA have provided a glimpse into the molecular dynamics of replication fork stalling. We report that a combination of density-based replisome enrichment by isolating proteins on nascent DNA (iPOND2) and label-free quantitative mass spectrometry (iPOND2-DRIPPER) substantially increases both replication factor yields and the dynamic range of protein quantification. Replication protein abundance in retrieved nascent DNA is elevated up to 300-fold over post-replicative controls, and recruitment of replication stress factors upon fork stalling is observed at similar levels...
May 2, 2024: Cell Reports