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LEDGF (p75) promotes DNA-end resection and homologous recombination

Nature Structural & Molecular Biology volume 19, pages 803–810 (2012)Cite this article

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Abstract

Lens epithelium–derived growth factor p75 splice variant (LEDGF) is a chromatin-binding protein known for its antiapoptotic activity and ability to direct human immunodeficiency virus into active transcription units. Here we show that LEDGF promotes the repair of DNA double-strand breaks (DSBs) by the homologous recombination repair pathway. Depletion of LEDGF impairs the recruitment of C-terminal binding protein interacting protein (CtIP) to DNA DSBs and the subsequent CtIP-dependent DNA-end resection. LEDGF is constitutively associated with chromatin through its Pro-Trp-Trp-Pro (PWWP) domain that binds preferentially to epigenetic methyl-lysine histone markers characteristic of active transcription units. LEDGF binds CtIP in a DNA damage–dependent manner, thereby enhancing its tethering to the active chromatin and facilitating its access to DNA DSBs. These data highlight the role of PWWP-domain proteins in DNA repair and provide a molecular explanation for the antiapoptotic and cancer cell survival–activities of LEDGF.

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Figure 1: LEDGF depletion sensitizes cells to ionizing radiation, camptothecin and mitomycin C.
Figure 2: LEDGF depletion inhibits DNA repair.
Figure 3: LEDGF facilitates homologous recombination in S- and G2-phase cells.
Figure 4: LEDGF promotes DNA-end resection.
Figure 5: Chromatin-bound LEDGF tethers CtIP to chromatin.
Figure 6: CtIP is tethered to LEDGF-decorated active chromatin markers.

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Acknowledgements

We thank A. Engelman (Dana-Farber Cancer Institute, Harvard Medical School) for providing us with Psip1-deficient cells, K.G. Henriksen for technical assistance and L. Vinner for discussions regarding HIV integration. This work was supported by the Danish Cancer Society and the Novo Nordisk Foundation (M.D., J.L., J.B. and M.J.), the Danish National Research Foundation and the John and Birthe Meyer Foundation (J.L., J.B. and M.J.), the Danish Medical Research Council, the Lundbeck Foundation and the European Commission FP7 (APO-SYS) (M.J.), and DDResponse, Biomedreg and INFLA-CARE (J.B).

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Author notes

  1. Lou Klitgaard Povlsen

    Present address: Present address: Novo Nordisk Foundation Center for Protein Research, Copenhagen University, Copenhagen, Denmark.,

Authors and Affiliations

  1. Cell Death and Metabolism, Danish Cancer Society Research Center, Copenhagen, Denmark

    Mads Daugaard, Annika Baude, Lou Klitgaard Povlsen, Nikolaj H T Petersen, Mikkel Rohde & Marja Jäättelä

  2. Center for Genotoxic Stress Research, Danish Cancer Society Research Center, Copenhagen, Denmark

    Mads Daugaard, Annika Baude, Lou Klitgaard Povlsen, Nikolaj H T Petersen, Claudia Lukas, Jiri Bartek, Jiri Lukas, Mikkel Rohde & Marja Jäättelä

  3. British Columbia Cancer Research Centre, University of British Columbia, Vancouver, British Columbia, Canada

    Mads Daugaard & Poul H B Sorensen

  4. Biotech Research and Innovation Centre, Copenhagen, Denmark.,

    Kasper Fugger, Halfdan Beck & Claus Storgaard Sørensen

  5. Chromosome Biology, Danish Cancer Society Research Center, Copenhagen, Denmark

    Claudia Lukas & Jiri Lukas

  6. Genome Integrity, Danish Cancer Society Research Center, Copenhagen, Denmark

    Jiri Bartek

  7. Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic

    Jiri Bartek

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Contributions

M.D. and M.J. designed the project and wrote the manuscript; M.D. and A.B. performed most of the experiments, supported by K.F., L.K.P., H.B., N.H.T.P. and M.R.; and C.S.S., C.L., J.L., J.B. and P.H.B.S. provided essential knowhow and support for the project and for preparing the manuscript.

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Correspondence to Mads Daugaard or Marja Jäättelä.

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Daugaard, M., Baude, A., Fugger, K. et al. LEDGF (p75) promotes DNA-end resection and homologous recombination. Nat Struct Mol Biol 19, 803–810 (2012). https://doi.org/10.1038/nsmb.2314

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