The end-joining factor Ku acts in the end-resection of double strand break-free arrested replication forks

Ana Teixeira-Silva, Anissia Ait Saada, Julien Hardy, Ismail Iraqui, Marina Charlotte Nocente, Karine Fréon and Sarah A. E. Lambert ()
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Ana Teixeira-Silva: PSL Research University, CNRS, UMR3348
Anissia Ait Saada: PSL Research University, CNRS, UMR3348
Julien Hardy: PSL Research University, CNRS, UMR3348
Ismail Iraqui: PSL Research University, CNRS, UMR3348
Marina Charlotte Nocente: PSL Research University, CNRS, UMR3348
Karine Fréon: PSL Research University, CNRS, UMR3348
Sarah A. E. Lambert: PSL Research University, CNRS, UMR3348

Nature Communications, 2017, vol. 8, issue 1, 1-14

Abstract: Abstract Replication requires homologous recombination (HR) to stabilize and restart terminally arrested forks. HR-mediated fork processing requires single stranded DNA (ssDNA) gaps and not necessarily double strand breaks. We used genetic and molecular assays to investigate fork-resection and restart at dysfunctional, unbroken forks in Schizosaccharomyces pombe. Here, we report that fork-resection is a two-step process regulated by the non-homologous end joining factor Ku. An initial resection mediated by MRN-Ctp1 removes Ku from terminally arrested forks, generating ~110 bp sized gaps obligatory for subsequent Exo1-mediated long-range resection and replication restart. The mere lack of Ku impacts the processing of arrested forks, leading to an extensive resection, a reduced recruitment of RPA and Rad51 and a slower fork-restart process. We propose that terminally arrested forks undergo fork reversal, providing a single DNA end for Ku binding. We uncover a role for Ku in regulating end-resection of unbroken forks and in fine-tuning HR-mediated replication restart.

Date: 2017
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DOI: 10.1038/s41467-017-02144-5

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