Replication dynamics of recombination-dependent replication forks

Naiman, Karel, Campillo-Funollet, Eduard, Watson, Adam T, Budden, Alice, Miyabe, Izumi and Carr, Antony M (2021) Replication dynamics of recombination-dependent replication forks. Nature Communications, 12. a923 1-11. ISSN 2041-1723

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Replication forks restarted by homologous recombination are error prone and replicate both strands semi-conservatively using Pol δ. Here, we use polymerase usage sequencing to visualize in vivo replication dynamics of HR-restarted forks at an S. pombe replication barrier, RTS1, and model replication by Monte Carlo simulation. We show that HR-restarted forks synthesise both strands with Pol δ for up to 30 kb without maturing to a δ/ε configuration and that Pol α is not used significantly on either strand, suggesting the lagging strand template remains as a gap that is filled in by Pol δ later. We further demonstrate that HR-restarted forks progress uninterrupted through a fork barrier that arrests canonical forks. Finally, by manipulating lagging strand resection during HR-restart by deleting pku70, we show that the leading strand initiates replication at the same position, signifying the stability of the 3′ single strand in the context of increased resection.

Item Type: Article
Keywords: Replication, DNA repair
Schools and Departments: School of Life Sciences > Sussex Centre for Genome Damage and Stability
Research Centres and Groups: Genome Damage and Stability Centre
Subjects: Q Science > QR Microbiology > QR0001 General
Depositing User: Antony Carr
Date Deposited: 19 Jan 2021 13:17
Last Modified: 16 Feb 2021 15:00

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Project NameSussex Project NumberFunderFunder Ref
Replication arrest, restart and genome instabilityG1829WELLCOME TRUST110047/Z/15/Z