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

• Rémy Bétous ^[1] ; Marie-Jeanne Pillaire ^[1] ; Laura Pierini ^[1] ; Siem van der Laan ^[4] ; Bénédicte Recolin ^[4] ; Emma Ohl-Séguy ^[1] ; Caixia Guo ^[2] ; Naoko Niimi ^[3] ; Petr Grúz ^[3] ; Takehiko Nohmi ^[3] ; Errol Friedberg ^[2] ; Christophe Cazaux ^[1] ; Domenico Maiorano ^[4] ; Jean-Sébastien Hoffmann ^[1]
  1. [1] Université de Toulouse

     Université de Toulouse

     Arrondissement de Toulouse, Francia

     
  2. [2] University of Texas Southwestern Medical Center

     University of Texas Southwestern Medical Center

     Estados Unidos

     
  3. [3] National Institute of Health Sciences

     National Institute of Health Sciences

     Japón

     
  4. [4] Institut de Génétique Humaine (IGH), CNRS—UPR 1142, Montpellier Cedex 5, France

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• Localización: EMBO journal: European Molecular Biology Organization, ISSN 0261-4189, Vol. 32, Nº. 15, 2013, págs. 2172-2185
• Idioma: inglés
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  • 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.