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The completion of genome duplication during the cell cycle is threatened by the presence of replication fork barriers (RFBs). Following collision with a RFB, replication proteins can dissociate from the stalled fork (fork collapse) rendering it incapable of further DNA synthesis unless recombination intervenes to restart replication. We use time-lapse microscopy and genetic assays to show that recombination is initiated within ∼ 10 min of replication fork blockage at a site-specific barrier in fission yeast, leading to a restarted fork within ∼ 60 min, which is only prevented/curtailed by the arrival of the opposing replication fork. The restarted fork is susceptible to further collapse causing hyper-recombination downstream of the barrier. Surprisingly, in our system fork restart is unnecessary for maintaining cell viability. Seemingly, the risk of failing to complete replication prior to mitosis is sufficient to warrant the induction of recombination even though it can cause deleterious genetic change.

Original publication

DOI

10.7554/eLife.04539

Type

Journal article

Journal

Elife

Publication Date

25/03/2015

Volume

4

Keywords

DNA replication, S. pombe, chromosomes, genes, homologous recombination, replication fork barrier, replication restart, Bacterial Proteins, Cell Division, DNA Helicases, DNA Replication, DNA, Fungal, DNA-Binding Proteins, Gene Expression Regulation, Fungal, Genes, Reporter, Luminescent Proteins, Microscopy, Fluorescence, Mitosis, Protein Phosphatase 2, Rad51 Recombinase, Recombination, Genetic, Schizosaccharomyces, Schizosaccharomyces pombe Proteins, Time-Lapse Imaging