DNA polymerase κ-dependent DNA synthesis at stalled replication forks is important for CHK1 activation

Rémy Bétous, Marie Jeanne Pillaire, Laura Pierini, Siem Van Der Laan, Bénédicte Recolin, Emma Ohl-Séguy, Caixia Guo, Naoko Niimi, Petr Grúz, Takehiko Nohmi, Errol Friedberg, Christophe Cazaux, Domenico Maiorano, Jean Sébastien Hoffmann

Research output: Contribution to journalArticlepeer-review

42 Scopus citations

Abstract

Formation of primed single-stranded DNA at stalled replication forks triggers activation of the replication checkpoint signalling cascade resulting in the ATR-mediated phosphorylation of the Chk1 protein kinase, thus preventing genomic instability. By using siRNA-mediated depletion in human cells and immunodepletion and reconstitution experiments in Xenopus egg extracts, we report that the Y-family translesion (TLS) DNA polymerase kappa (Pol κ) contributes to the replication checkpoint response and is required for recovery after replication stress. We found that Pol κ is implicated in the synthesis of short DNA intermediates at stalled forks, facilitating the recruitment of the 9-1-1 checkpoint clamp. Furthermore, we show that Pol κ interacts with the Rad9 subunit of the 9-1-1 complex. Finally, we show that this novel checkpoint function of Pol κ is required for the maintenance of genomic stability and cell proliferation in unstressed human cells.

Original languageEnglish (US)
Pages (from-to)2172-2185
Number of pages14
JournalEMBO Journal
Volume32
Issue number15
DOIs
StatePublished - Jul 31 2013

Keywords

  • DNA polymerase κ
  • genetic instability
  • replication checkpoint
  • replication stress

ASJC Scopus subject areas

  • Neuroscience(all)
  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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