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DVC1 (C1orf124) is a DNA damage–targeting p97 adaptor that promotes ubiquitin-dependent responses to replication blocks

Nature Structural & Molecular Biology volume 19, pages 10841092 (2012) | Download Citation

Abstract

Ubiquitin-mediated processes orchestrate critical DNA-damage signaling and repair pathways. We identify human DVC1 (C1orf124; Spartan) as a cell cycle–regulated anaphase-promoting complex (APC) substrate that accumulates at stalled replication forks. DVC1 recruitment to sites of replication stress requires its ubiquitin-binding UBZ domain and PCNA-binding PIP box motif but is independent of RAD18-mediated PCNA monoubiquitylation. Via a conserved SHP box, DVC1 recruits the ubiquitin-selective chaperone p97 to blocked replication forks, which may facilitate p97-dependent removal of translesion synthesis (TLS) DNA polymerase η (Pol η) from monoubiquitylated PCNA. DVC1 knockdown enhances UV light–induced mutagenesis, and depletion of human DVC1 or the Caenorhabditis elegans ortholog DVC-1 causes hypersensitivity to replication stress–inducing agents. Our findings establish DVC1 as a DNA damage–targeting p97 adaptor that protects cells from deleterious consequences of replication blocks and suggest an important role of p97 in ubiquitin-dependent regulation of TLS.

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Acknowledgements

We thank K. Ramadan (University of Zurich), A. Lehmann (University of Sussex), R. Pepperkok and J. Simpson (European Molecular Biology Laboratory, Heidelberg), G. Alexandru (University of Dundee, UK), S. Fang (University of Maryland), M. Seidman (US National Institute on Aging), and members of the Caenorhabditis Genetics Center for providing reagents and worm strains, C. Doil and P. Menard for help with the initial screen for factors accumulating at laser-induced DNA damage, and J. Rouse and I. Dikic for communicating results before publication. This work was supported by grants from the Novo Nordisk Foundation, Danish Medical Research Council, the Danish Cancer Society, the Lundbeck Foundation, the Danish National Research Foundation and the European Research Council (Starting grant to R.P.).

Author information

Author notes

    • Anna Mosbech
    •  & Ian Gibbs-Seymour

    These authors contributed equally to this work.

Affiliations

  1. Ubiquitin Signaling Group, Department of Disease Biology, Novo Nordisk Foundation Center for Protein Research, Faculty of Health Sciences, University of Copenhagen, Copenhagen, Denmark.

    • Anna Mosbech
    • , Ian Gibbs-Seymour
    • , Tina Thorslund
    • , Lou Povlsen
    • , Stine Smedegaard
    • , Simon Bekker-Jensen
    •  & Niels Mailand
  2. Biotech Research and Innovation Center, University of Copenhagen, Copenhagen, Denmark.

    • Konstantinos Kagias
    •  & Roger Pocock
  3. Department of Proteomics, Novo Nordisk Foundation Center for Protein Research, Faculty of Health Sciences, University of Copenhagen, Copenhagen, Denmark.

    • Petra Beli
    •  & Chunaram Choudhary
  4. Department of Biology, University of Copenhagen, Copenhagen, Denmark.

    • Sofie Vincents Nielsen
    •  & Rasmus Hartmann-Petersen
  5. Mitotic Mechanisms and Regulation, Department of Disease Biology, Novo Nordisk Foundation Center for Protein Research, Faculty of Health Sciences, University of Copenhagen, Copenhagen, Denmark.

    • Garry Sedgwick
  6. Centre for Genotoxic Stress Research and Chromosome Biology Unit, Danish Cancer Society Research Center, Copenhagen, Denmark.

    • Claudia Lukas
    •  & Jiri Lukas

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Contributions

A.M., I.G.-S., K.K. and T.T. performed most of the experiments, supported by L.P., S.V.N., S.B.-J. and G.S. S.S. provided stable U2OS GFP–Pol η cells. P.B. and C.C. carried out MS experiments and analyzed data from MS experiments. C.L. and J.L. provided essential information and support for initiation of the project. S.B.-J. and N.M. supervised the project and analyzed data, with participation by R.H.-P. and R.P.; N.M. wrote the paper.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Niels Mailand.

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DOI

https://doi.org/10.1038/nsmb.2395

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