Journal
NATURE STRUCTURAL & MOLECULAR BIOLOGY
Volume 19, Issue 1, Pages 17-U30Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nsmb.2177
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Funding
- Cancer Research UK
- European Research Council (ERC) [206281]
- Lister Institute of Preventive Medicine
- European Molecular Biology Organization (EMBO)
- European Research Council (ERC) [206281] Funding Source: European Research Council (ERC)
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In higher eukaryotes, the dynamics of replisome components during fork collapse and restart are poorly understood. Here we have reconstituted replication fork collapse and restart by inducing single-strand DNA lesions that create a double-strand break in one of the replicated sister chromatids after fork passage. We found that, upon fork collapse, the active CDC45-MCM-GINS (CMG) helicase complex loses its GINS subunit. A functional replisome is restored by the reloading of GINS and polymerase epsilon onto DNA in a fashion that is dependent on RAD51 and MRE11 but independent of replication origin assembly and firing. PCNA mutant alleles defective in break-induced replication (BIR) are unable to support restoration of replisome integrity. These results show that, in higher eukaryotes, replisomes are partially dismantled after fork collapse and fully re-established by a recombination-mediated process.
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