DNA replication is highly resilient and persistent under the challenge of mild replication stress

Summary

Mitotic DNA synthesis (MiDAS) has been proposed to restart DNA synthesis during mitosis because of replication fork stalling in late interphase caused by mild replication stress (RS). Contrary to this proposal, we find that cells exposed to mild RS in fact maintain continued DNA replication throughout G2 and during G2-M transition in RAD51- and RAD52-dependent manners. Persistent DNA synthesis is necessary to resolve replication intermediates accumulated in G2 and disengage an ATR-imposed block to mitotic entry. Because of its continual nature, DNA synthesis at very late replication sites can overlap with chromosome condensation, generating the phenomenon of mitotic DNA synthesis. Unexpectedly, we find that the commonly used CDK1 inhibitor RO3306 interferes with replication to preclude detection of G2 DNA synthesis, leading to the impression of a mitosis-driven response. Our study reveals the importance of persistent DNA replication and checkpoint control to lessen the risk for severe genome under-replication under mild RS.

Graphical abstract

Highlights

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  DNA synthesis persists during G2-M transition to counteract replication stress (RS)

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  RAD51/RAD52-mediated HR pathways facilitate the continuation of G2-M DNA synthesis

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  Continued G2 DNA synthesis relieves RS-induced G2/M checkpoint for mitotic entry

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  RO3306, but not CDK1 inhibition, non-specifically interferes with DNA synthesis

Abstract

Contrary to the MiDAS model, in which stressed replication forks fail to proceed in late interphase unless mitosis is initiated, Mocanu et al. reveal that cells maintain continued DNA synthesis from S-phase to early mitosis to minimize genome under-replication. This response is also essential to lessen persistent ATR-mediated G2/M checkpoint activation.