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.).
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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.
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Mosbech, A., Gibbs-Seymour, I., Kagias, K. et al. DVC1 (C1orf124) is a DNA damage–targeting p97 adaptor that promotes ubiquitin-dependent responses to replication blocks. Nat Struct Mol Biol 19, 1084–1092 (2012). https://doi.org/10.1038/nsmb.2395
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DOI: https://doi.org/10.1038/nsmb.2395