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The cell-cycle checkpoint kinase Chk1 is required for mammalian homologous recombination repair

Nature Cell Biology volume 7, pages 195–201 (2005)Cite this article

Abstract

The essential checkpoint kinase Chk1 is required for cell-cycle delays after DNA damage or blocked DNA replication1,2. However, it is unclear whether Chk1 is involved in the repair of damaged DNA. Here we establish that Chk1 is a key regulator of genome maintenance by the homologous recombination repair (HRR) system. Abrogation of Chk1 function with small interfering RNA or chemical antagonists inhibits HRR, leading to persistent unrepaired DNA double-strand breaks (DSBs) and cell death after replication inhibition with hydroxyurea or DNA-damage caused by camptothecin. After hydroxyurea treatment, the essential recombination repair protein RAD51 is recruited to DNA repair foci performing a vital role in correct HRR3,4. We demonstrate that Chk1 interacts with RAD51, and that RAD51 is phosphorylated on Thr 309 in a Chk1-dependent manner. Consistent with a functional interplay between Chk1 and RAD51, Chk1-depleted cells failed to form RAD51 nuclear foci after exposure to hydroxyurea, and cells expressing a phosphorylation-deficient mutant RAD51T309A were hypersensitive to hydroxyurea. These results highlight a crucial role for the Chk1 signalling pathway in protecting cells against lethal DNA lesions through regulation of HRR.

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Figure 1: A Chk1 signal to homologous recombination is required for survival after DNA damage and replication arrest.
Figure 2: Inhibition of Chk1 leads to increased accumulation of replication-associated DSBs and failure to repair DSBs after HU treatment.
Figure 3: Chk1 is required for RAD51 foci formation after hydroxyurea treatment.
Figure 4: Chk1 directly regulates RAD51.

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Acknowledgements

We thank M. Jasin for the SCneo and pCMV3xnlsI-SceI vectors, L. Thompson for cell lines and the Danish Cancer Society, the Novo Nordisk Foundation, the European Union, the Danish Medical Research Council, Dansk Kraeftforskningsfond, the Swedish Cancer Society, the Swedish Research Council, the Biological & Biotechnological Sciences Research Council and Yorkshire Cancer Research for grant support.

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Author notes

  1. Claus Storgaard Sørensen

    Present address: The Biotech Research and Innovation Centre (BRIC), Fruebjergvej 3, DK-2100, Copenhagen, Denmark

  2. Claus Storgaard Sørensen and Lasse Tengbjerg Hansen: These authors contributed equally to this work.

Authors and Affiliations

  1. Danish Cancer Society, Institute of Cancer Biology, Copenhagen, DK-2100, Denmark

    Claus Storgaard Sørensen, Randi G. Syljuåsen & Jiri Bartek

  2. Institute of Molecular Pathology, University of Copenhagen, Copenhagen, DK-2100, Denmark

    Lasse Tengbjerg Hansen

  3. The Institute for Cancer Studies, University of Sheffield, Sheffield, S10 2RX, UK

    Jaroslaw Dziegielewski & Thomas Helleday

  4. Department of Genetics Microbiology and Toxicology, Stockholm University, Stockholm, S-106 91, Sweden

    Cecilia Lundin & Thomas Helleday

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Correspondence to Jiri Bartek or Thomas Helleday.

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Sørensen, C., Hansen, L., Dziegielewski, J. et al. The cell-cycle checkpoint kinase Chk1 is required for mammalian homologous recombination repair. Nat Cell Biol 7, 195–201 (2005). https://doi.org/10.1038/ncb1212

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