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Regulation of Mus81–Eme1 Holliday junction resolvase in response to DNA damage

Nature Structural & Molecular Biology volume 20, pages 598–603 (2013)Cite this article

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Abstract

Structure-specific DNA endonucleases have critical roles during DNA replication, repair and recombination, yet they also have the potential for causing genome instability. Controlling these enzymes may be essential to ensure efficient processing of ad hoc substrates and to prevent random, unscheduled processing of other DNA structures, but it is unknown whether structure-specific endonucleases are regulated in response to DNA damage. Here, we uncover DNA damage–induced activation of Mus81–Eme1 Holliday junction resolvase in fission yeast. This new regulation requires both Cdc2CDK1- and Rad3ATR-dependent phosphorylation of Eme1. Mus81–Eme1 activation prevents gross chromosomal rearrangements in cells lacking the BLM-related DNA helicase Rqh1. We propose that linking Mus81–Eme1 DNA damage–induced activation to cell-cycle progression ensures efficient resolution of Holliday junctions that escape dissolution by Rqh1–TopIII while preventing unnecessary DNA cleavages.

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Figure 1: Phosphorylation of Eme1 and hyperactivation of Mus81–Eme1 in response to DNA damage.
Figure 2: Eme1 is phosphorylated by the DNA-damage checkpoint.
Figure 3: Eme1 is phosphorylated in a Cdc2CDK1-dependent manner.
Figure 4: Cdc2CDK1-dependent phosphorylation primes Eme1 for DNA damage–dependent hyperactivation of Mus81–Eme1.
Figure 5: Hyperactivation of Mus81–Eme1 is critical in the absence of Rqh1BLM.
Figure 6: DNA damage–induced phosphorylation of Eme1 prevents gross chromosomal rearrangements in the absence of Rqh1BLM.

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Acknowledgements

We thank M. Grenon, N. Rhind, C. Chahwan, N. Boddy, J.H. Guervilly and G. Almouzni for critical reading of the manuscript and stimulating discussions throughout the course of this study and L. Brossy for technical help. Many thanks to V. Simanis (École Polytechnique Fédérale de Lausanne, Lausanne, Switzerland) for kindly providing the cdc2as strain and to Y. Denis for technical help with the CGH analysis. P.-H.L.G. acknowledges financial support from the Fondation Arc pour la Recherche sur le Cancer (SF1052), Centre National de la Recherche Scientifique (ATIP program) and Agence Nationale de la Recherche (ANR-10-BLAN-1512-01) of France. B.L. was supported by a grant from the Agence Nationale de la Recherche (ANR-10-BLAN-1606-03). Work in the laboratory of P.R. was supported by the US National Institutes of Health (RO1 GM59447).

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Author notes

  1. James A Wohlschlegel

    Present address: Current address: Department of Biological Chemistry, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, California, USA.,

  2. Pierre-Marie Dehé and Stéphane Coulon: These authors contributed equally to this work.

Authors and Affiliations

  1. Centre National de la Recherche Scientifique, Unité Mixte de Recherche 7258, Inserm-Unité 1068, Centre de Recherche en Cancérologie de Marseille, Institut Paoli-Calmettes, France

    Pierre-Marie Dehé, Stéphane Coulon, Sarah Scaglione, Arato Takedachi, Bertrand Llorente & Pierre-Henri L Gaillard

  2. Aix-Marseille Université, F-13284, Marseille, France

    Pierre-Marie Dehé, Stéphane Coulon, Sarah Scaglione, Arato Takedachi, Bertrand Llorente & Pierre-Henri L Gaillard

  3. Department of Molecular Biology, Scripps Research Institute, La Jolla, California, USA

    Paul Shanahan & Paul Russell

  4. Department of Chemical Physiology, Scripps Research Institute, La Jolla, California, USA

    James A Wohlschlegel & John R Yates III

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Contributions

P.-M.D., S.C., S.S., A.T., P.S., J.A.W. and P.-H.L.G. designed and performed experiments. P.-H.L.G., P.R., B.L. and J.R.Y. designed experiments and supervised the work, and P.-H.L.G. wrote the paper. Experiments in Figures 1, 2 and 4 were performed by P.-M.D. and S.S. with initial input from P.-H.L.G., P.S. and S.C. in strain engineering. Experiments in Figure 3 were all performed by P.-M.D. with initial input from P.-H.L.G. and P.S. in strain engineering for Figure 3a,b. Experiments in Figure 5a,b were performed by P.-M.D., in Figure 5c,d by P.-H.L.G. and in Figure 6a,b by S.C.

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Correspondence to Pierre-Henri L Gaillard.

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Dehé, PM., Coulon, S., Scaglione, S. et al. Regulation of Mus81–Eme1 Holliday junction resolvase in response to DNA damage. Nat Struct Mol Biol 20, 598–603 (2013). https://doi.org/10.1038/nsmb.2550

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