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PERK inhibits DNA replication during the Unfolded Protein Response via Claspin and Chk1

Oncogene volume 36, pages 678–686 (2017)Cite this article

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Abstract

Stresses such as hypoxia, nutrient deprivation and acidification disturb protein folding in the endoplasmic reticulum (ER) and activate the Unfolded Protein Response (UPR) to trigger adaptive responses through the effectors, PERK, IRE1 and ATF6. Most of these responses relate to ER homoeostasis; however, here we show that the PERK branch of the UPR also controls DNA replication. Treatment of cells with the non-genotoxic UPR agonist thapsigargin led to a rapid inhibition of DNA synthesis that was attributable to a combination of DNA replication fork slowing and reduced replication origin firing. DNA synthesis inhibition was dependent on the UPR effector PERK and was associated with phosphorylation of the checkpoint adaptor protein Claspin and activation of the Chk1 effector kinase, both of which occurred in the absence of detectable DNA damage. Remarkably, thapsigargin did not inhibit bulk DNA synthesis or activate Chk1 in cells depleted of Claspin, or when Chk1 was depleted or subject to chemical inhibition. In each case thapsigargin-resistant DNA synthesis was due to an increase in replication origin firing that compensated for reduced fork progression. Taken together, our results unveil a new aspect of PERK function and previously unknown roles for Claspin and Chk1 as negative regulators of DNA replication in the absence of genotoxic stress. Because tumour cells proliferate in suboptimal environments, and frequently show evidence of UPR activation, this pathway could modulate the response to DNA replication-targeted chemotherapies.

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Acknowledgements

This work was supported by grants from the Spanish Ministry of Economy and Competitiveness (SAF2013-49149-R, BFU2014-51672-REDC to RF) and Fundacion CajaCanarias (AP2015/008 to RF), the IMBRAIN Project (FP7-REGPOT-2012-CT2012-31637-IMBRAIN to DAG) funded by EU FP7 and Gobierno de Canarias, and World Wide Cancer Research (WWCR; Project grant no. 12-0149 to DAG).

Author contributions

EC, SH, SK, MD, DK and MS performed experiments and data analysis. EC, SH, RF and DAG designed experiments and prepared the manuscript.

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Authors and Affiliations

  1. Unidad de Investigación, Hospital Universitario de Canarias, Instituto de Tecnologías Biomédicas, Ofra s/n, La Cuesta, La Laguna, Tenerife, Spain

    E Cabrera, S Hernández-Pérez & R Freire

  2. Institut Curie, Centre de Recherche, CNRS-UMR 3244, Paris Cedex 05, France

    S Koundrioukoff & M Debatisse

  3. Sorbonne Universités, UPMC Univ Paris 06, Paris, France

    S Koundrioukoff & M Debatisse

  4. CNRS-UMR 3244, Paris, France

    S Koundrioukoff & M Debatisse

  5. Department of Molecular Biology and Genetics, Weill Institute for Cell and Molecular Biology, Cornell University, Ithaca, NY, USA

    D Kim & M B Smolka

  6. Instituto de Tecnologías Biomédicas, Centro de Investigaciones Biomédicas de Canarias, Facultad de Medicina, Campus Ciencias de la Salud, Universidad de La Laguna, La Laguna, Tenerife, Spain

    D A Gillespie

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  1. E Cabrera
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  2. S Hernández-Pérez
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  8. D A Gillespie
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Correspondence to R Freire or D A Gillespie.

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Cabrera, E., Hernández-Pérez, S., Koundrioukoff, S. et al. PERK inhibits DNA replication during the Unfolded Protein Response via Claspin and Chk1. Oncogene 36, 678–686 (2017). https://doi.org/10.1038/onc.2016.239

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