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

Author

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  • Ana Teixeira-Silva

    (PSL Research University, CNRS, UMR3348
    University Paris Sud, Paris-Saclay University, CNRS, UMR3348)

  • Anissia Ait Saada

    (PSL Research University, CNRS, UMR3348
    University Paris Sud, Paris-Saclay University, CNRS, UMR3348)

  • Julien Hardy

    (PSL Research University, CNRS, UMR3348
    University Paris Sud, Paris-Saclay University, CNRS, UMR3348)

  • Ismail Iraqui

    (PSL Research University, CNRS, UMR3348
    University Paris Sud, Paris-Saclay University, CNRS, UMR3348)

  • Marina Charlotte Nocente

    (PSL Research University, CNRS, UMR3348
    University Paris Sud, Paris-Saclay University, CNRS, UMR3348)

  • Karine Fréon

    (PSL Research University, CNRS, UMR3348
    University Paris Sud, Paris-Saclay University, CNRS, UMR3348)

  • Sarah A. E. Lambert

    (PSL Research University, CNRS, UMR3348
    University Paris Sud, Paris-Saclay University, CNRS, UMR3348)

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.

Suggested Citation

  • Ana Teixeira-Silva & Anissia Ait Saada & Julien Hardy & Ismail Iraqui & Marina Charlotte Nocente & Karine Fréon & Sarah A. E. Lambert, 2017. "The end-joining factor Ku acts in the end-resection of double strand break-free arrested replication forks," Nature Communications, Nature, vol. 8(1), pages 1-14, December.
    • Handle: RePEc:nat:natcom:v:8:y:2017:i:1:d:10.1038_s41467-017-02144-5
    DOI: 10.1038/s41467-017-02144-5
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