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HERC2 coordinates ubiquitin-dependent assembly of DNA repair factors on damaged chromosomes

Nature Cell Biology volume 12, pages 80–86 (2010)Cite this article

A Corrigendum to this article was published on 01 April 2010

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Abstract

Regulatory ubiquitylation is emerging as an important mechanism to protect genome integrity in cells exposed to DNA damage1,2,3,4,5,6,7,8,9. However, the spectrum of known ubiquitin regulators of the DNA damage response (DDR) is limited and their functional interplay is poorly understood. Here, we identify HERC2 as a factor that regulates ubiquitin-dependent retention of repair proteins on damaged chromosomes. In response to ionising radiation (IR), HERC2 forms a complex with RNF8, a ubiquitin ligase involved in the DDR3,4,5,6. The HERC2–RNF8 interaction requires IR-inducible phosphorylation of HERC2 at Thr 4827, which in turn binds to the forkhead-associated (FHA) domain of RNF8. Mechanistically, we provide evidence that HERC2 facilitates assembly of the ubiquitin-conjugating enzyme Ubc13 with RNF8, thereby promoting DNA damage-induced formation of Lys 63-linked ubiquitin chains. We also show that HERC2 interacts with, and maintains the levels of, RNF168, another ubiquitin ligase operating downstream of RNF8 (Refs 7, 8). Consequently, knockdown of HERC2 abrogates ubiquitin-dependent retention of repair factors such as 53BP1, RAP80 and BRCA1. Together with the increased radiosensitivity of HERC2-depleted cells, these results uncover a regulatory layer in the orchestration of protein interactions on damaged chromosomes and they underscore the role of ubiquitin-mediated signalling in genome maintenance.

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Figure 1: HERC2 associates with RNF8 and accumulates at DSB sites in an RNF8-dependent manner.
Figure 2: Phosphorylation of HERC2 on Thr 4827 promotes binding to the FHA domain of RNF8.
Figure 3: HERC2 is required for RNF8-mediated ubiquitylation at DSB sites and for recruitment of DNA repair factors.
Figure 4: HERC2 functions as an assembly factor for the RNF8–Ubc13 complex to promote the RNF8-mediated DDR.
Figure 5: HERC2 binds and stabilizes RNF168.

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Change history

  • 10 March 2010

    In the version of this letter initially published online, an author Claudia Lukas, was omitted. This error has been corrected in both the HTML and PDF versions of the letter.

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Acknowledgements

We thank Z. Ronai and D. Durocher for reagents, and J Chen for sharing unpublished data. This work was supported by grants from the Danish Cancer Society, Danish National Research Foundation, European Commission (integrated projects GENICA and DNA Repair), Lundbeck Foundation, Danish Research Council and the John and Birthe Meyer Foundation.

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Author notes

  1. Simon Bekker-Jensen, Jannie Rendtlew Danielsen & Niels Mailand

    Present address: Current address: The Novo Nordisk Foundation Center for Protein Research, Faculty of Health Sciences, University of Copenhagen, Blegdamsvej 3B, DK-2200, Copenhagen, Denmark.,

  2. Simon Bekker-Jensen and Jannie Rendtlew Danielsen: These authors contributed equally to this work.

Authors and Affiliations

  1. Centre for Genotoxic Stress Research, Strandboulevarden 49, Copenhagen, DK-2100, Denmark

    Simon Bekker-Jensen, Jannie Rendtlew Danielsen, Kasper Fugger, Annika Nerstedt, Claudia Lukas, Jiri Bartek, Jiri Lukas & Niels Mailand

  2. Department of Proteomics in Cancer, Institute of Cancer Biology, Danish Cancer Society, Strandboulevarden 49, Copenhagen, DK-2100, Denmark

    Irina Gromova

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Contributions

N.M., S.B.-J., J.B. and J.L. designed the project and wrote the paper. N.M., S.B.-J. and J.R.D. performed most of the biochemical, cell biology and phosphorylation/clonogenic survival experiments, respectively. C.L. provided the in situ RNF168 data, I.G. performed the mass spectrometry analysis, A.N. generated the HERC2 constructs and K.F. contributed to the analysis of HERC2 complexes. All authors participated in interpreting the results.

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Correspondence to Jiri Bartek or Jiri Lukas.

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The authors declare no competing financial interests.

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Bekker-Jensen, S., Danielsen, J., Fugger, K. et al. HERC2 coordinates ubiquitin-dependent assembly of DNA repair factors on damaged chromosomes. Nat Cell Biol 12, 80–86 (2010). https://doi.org/10.1038/ncb2008

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