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Published Online November 15, 2007
Science DOI: 10.1126/science.1150034

Reports

Submitted on September 4, 2007
Accepted on November 5, 2007

Orchestration of the DNA-Damage Response by the RNF8 Ubiquitin Ligase

Nadine K. Kolas 1{dagger}, J. Ross Chapman 2{dagger}, Shinichiro Nakada 1{dagger}, Jarkko Ylanko 3, Richard Chahwan 2, Frédéric D. Sweeney 3, Stephanie Panier 1, Megan Mendez 1, Jan Wildenhain 1, Timothy M. Thomson 4, Laurence Pelletier 3, Stephen P. Jackson 2*, Daniel Durocher 3*

1 Samuel Lunenfeld Research Institute, Mount Sinai Hospital, 600 University Avenue, Toronto, M5G 1X5, Ontario, Canada.
2 The Wellcome Trust and Cancer Research UK Gurdon Institute, and the Department of Zoology, University of Cambridge, Tennis Court Road, Cambridge CB2 1QN, UK.
3 Samuel Lunenfeld Research Institute, Mount Sinai Hospital, 600 University Avenue, Toronto, M5G 1X5, Ontario, Canada.; Department of Molecular Genetics, University of Toronto, Toronto, Ontario, Canada.
4 Department of Molecular and Cellular Biology, Instituto de Biología Molecular de Barcelona c. Jordi Girona 18-2608034 Barcelona, Spain.

* To whom correspondence should be addressed.
Stephen P. Jackson , E-mail: durocher{at}mshri.on.ca
Daniel Durocher , E-mail: durocher{at}mshri.on.ca

{dagger}These authors contributed equally to this work.

Cells respond to DNA double-strand breaks (DSBs) by recruiting factors such as the DNA-damage mediator protein MDC1, p53-binding protein 1 (53BP1), and the breast cancer susceptibility protein BRCA1 to sites of damaged DNA. Here, we reveal that the ubiquitin ligase RNF8 mediates 53BP1 and BRCA1 focal accumulation at sites of DNA lesions. Moreover, we establish that MDC1 recruits RNF8 via phosphodependent interactions between the RNF8 forkhead-associated (FHA) domain and motifs in MDC1 that are phosphorylated by the DNA-damage activated protein kinase ATM. We also show that depletion of the E2 enzyme UBC13 impairs 53BP1 recruitment to sites of damage, suggesting that it cooperates with RNF8. Finally, we reveal that RNF8 promotes the G2/M DNA damage checkpoint and resistance to ionizing radiation. These results demonstrate how the DNA-damage response is orchestrated by both ATM-dependent phosphorylation of MDC1 and RNF8-mediated ubiquitination.


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