DNA polymerase κ‐dependent DNA synthesis at stalled replication forks is important for CHK1 activation |
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Authors: | Rémy Bétous Marie‐Jeanne Pillaire Laura Pierini Siem van der Laan Emma Ohl‐Séguy Caixia Guo Naoko Niimi Petr Grúz Takehiko Nohmi Errol Friedberg Christophe Cazaux Domenico Maiorano Jean‐Sébastien Hoffmann |
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Institution: | 1. Equipe Labellisée La Ligue Contre le Cancer 2013, INSERM UMR 1037, CNRS ERL 505294, CRCT (Cancer Research Center of Toulouse), , Toulouse, France;2. University of Toulouse;3. UPS, , Toulouse, France;4. Institut de Génétique Humaine (IGH), CNRS—UPR 1142, , Montpellier Cedex 5, France;5. Department of Pathology, The University of Texas Southwestern Medical Center, , Dallas, TX, USA;6. Division of Genetics and Mutagenesis, National Institute of Health Sciences, , Tokyo, Japan |
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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. |
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Keywords: | DNA polymerase κ genetic instability replication checkpoint replication stress |
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