Stochastic Endogenous Replication Stress Causes ATR-Triggered Fluctuations in CDK2 Activity that Dynamically Adjust Global DNA Synthesis Rates

Faithful DNA replication is challenged by stalling of replication forks during S phase. Replication stress is further increased in cancer cells or in response to genotoxic insults. Using live single-cell image analysis, we found that CDK2 activity fluctuates throughout an unperturbed S phase. We show that CDK2 fluctuations result from transient ATR signals triggered by stochastic replication stress events. In turn, fluctuating endogenous CDK2 activity causes corresponding decreases and increases in DNA synthesis rates, linking changes in stochastic replication stress to fluctuating global DNA replication rates throughout S phase. Moreover, cells that re-enter the cell cycle after mitogen stimulation have increased CDK2 fluctuations and prolonged S phase resulting from increased replication stress-induced CDK2 suppression. Thus, our study reveals a dynamic control principle for DNA replication whereby CDK2 activity is suppressed and fluctuates throughout S phase to continually adjust global DNA synthesis rates in response to recurring stochastic replication stress events. [Display omitted] •Live single-cell analysis reveals fluctuating CDK2 activity in S phase•Fluctuating CDK2 activity is caused by endogenous replication stress•CDK2 activity fluctuates to adjust global DNA synthesis rate•Serum-starved cells exit quiescence with more replication stress and longer S phase Live single-cell microscopy reveals a control principal that helps maintain proper duplication of genetic material. Upon inevitable DNA replication stress during S phase, cells signal through ATR to attenuate CDK2 activity, which then decreases global DNA synthesis rate. This feedback enables dynamic modulation of S-phase progression.