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Nat Struct Mol Biol. 2011 Dec 4;19(1):17-24. doi: 10.1038/nsmb.2177.

RAD51- and MRE11-dependent reassembly of uncoupled CMG helicase complex at collapsed replication forks.

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  • 1Genome Stability Unit, Clare Hall Laboratories, London Research Institute, South Mimms, Hertfordshire, UK.

Abstract

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 ɛ 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.

PMID:
22139015
[PubMed - indexed for MEDLINE]
PMCID:
PMC4306020
Free PMC Article
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