Letter | Published:

H4K20me0 marks post-replicative chromatin and recruits the TONSL–MMS22L DNA repair complex

Nature volume 534, pages 714718 (30 June 2016) | Download Citation

Abstract

After DNA replication, chromosomal processes including DNA repair and transcription take place in the context of sister chromatids. While cell cycle regulation can guide these processes globally, mechanisms to distinguish pre- and post-replicative states locally remain unknown. Here we reveal that new histones incorporated during DNA replication provide a signature of post-replicative chromatin, read by the human TONSL–MMS22L1,2,3,4 homologous recombination complex. We identify the TONSL ankyrin repeat domain (ARD) as a reader of histone H4 tails unmethylated at K20 (H4K20me0), which are specific to new histones incorporated during DNA replication and mark post-replicative chromatin until the G2/M phase of the cell cycle. Accordingly, TONSL–MMS22L binds new histones H3–H4 both before and after incorporation into nucleosomes, remaining on replicated chromatin until late G2/M. H4K20me0 recognition is required for TONSL–MMS22L binding to chromatin and accumulation at challenged replication forks and DNA lesions. Consequently, TONSL ARD mutants are toxic, compromising genome stability, cell viability and resistance to replication stress. Together, these data reveal a histone-reader-based mechanism for recognizing the post-replicative state, offering a new angle to understand DNA repair with the potential for targeted cancer therapy.

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Accessions

Primary accessions

Protein Data Bank

Data deposits

Coordinate and structure factors have been deposited in the Protein Data Bank under accession number 5JA4.

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Acknowledgements

We thank the beam staff at the synchrotrons at the Argonne National Laboratory (NE-CAT) for technical assistance. We thank J. Rouse, D. Durocher, G. Legube and C. Storgaard Sørensen for reagents, G. Montoya for assistance with circular dichroism, C. B. Strømme, A. Strandsby, K. Nakamura, S.-b. Lee and M. Hödl for help with experiments, and Y. Antoku for assistance with microscopy. We thank J. Lukas for comments on the manuscript and Z. Jasencakova for illustrations. G.S. was supported by European Commission Marie Curie ITN FP7 ‘aDDRess’. D.J.P. was supported in part by grants from the Leukemia and Lymphoma Society and the STARR foundation. A.G. is an EMBO Young Investigator and her research is supported by the European Research Council (ERC StG, no. 281765), the Danish National Research Foundation to the Center for Epigenetics (DNRF82), the Danish Cancer Society, the Danish Medical Research Council, the Novo Nordisk Foundation and the Lundbeck Foundation. A.I. is supported by the European Commission FP7 Network of Excellence EpiGeneSys (project 257082), the DFG Excellence Clusters CIPSM and SyNergy, as well as the DFG Collaborative Research Center 1064 (projects A3 and Z3). T.B. is supported by the Medical Research Council and the European Research Council (ERC StG, no. 309952).

Author information

Author notes

    • Giulia Saredi
    •  & Hongda Huang

    These authors contributed equally to this work.

Affiliations

  1. Biotech Research and Innovation Centre (BRIC) and Centre for Epigenetics, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen DK-2200, Denmark

    • Giulia Saredi
    • , Colin M. Hammond
    • , Constance Alabert
    • , Nazaret Reverón-Gómez
    •  & Anja Groth
  2. Structural Biology Program, Memorial Sloan-Kettering Cancer Center, New York, New York 10065, USA

    • Hongda Huang
    •  & Dinshaw J. Patel
  3. The Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, Copenhagen DK-2200, Denmark

    • Simon Bekker-Jensen
    •  & Niels Mailand
  4. Department of Molecular Biology, Biomedical Center and Center for Integrated Protein Science Munich, Ludwig-Maximilians University, 80336 Munich, Germany

    • Ignasi Forne
    •  & Axel Imhof
  5. MRC Clinical Sciences Centre (CSC) and Institute of Clinical Sciences (ICS), Faculty of Medicine, Imperial College London, Du Cane Road, London W12 0NN, UK

    • Benjamin M. Foster
    •  & Till Bartke
  6. Institute of Molecular Cancer Research, University of Zurich, Zurich CH-8057, Switzerland

    • Lucie Mlejnkova
    •  & Petr Cejka

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Contributions

G.S. and A.G. conceived and led the functional studies. H.H conceived and led the generation of cassette to crystallize the complex, and H.H. solved the structure and performed the ITC under the supervision of D.J.P. C.M.H. performed peptide pull-downs with ARD and recombinant nucleosomes. C.A. performed SET8 experiments and NCC. S.B.-J. and N.M. analysed recruitment to laser-induced DNA damage. N.R.-G. prepared histones for mass spectrometry and performed ChIP analysis, I.F. analysed histone modifications by mass spectrometry under the supervision of A.I. B.M.F and T.B. prepared modified recombinant nucleosomes. L.M. and P.C. prepared recombinant TONSL–MMS22L. G.S., H.H., D.J.P. and A.G. wrote the manuscript and all authors commented on the manuscript.

Competing interests

G.S., H.H., C.M.H., D.J.P. and A.G. are inventors on a filed patent application covering the discoveries presented in this manuscript.

Corresponding authors

Correspondence to Dinshaw J. Patel or Anja Groth.

Reviewer Information Nature thanks T. Kutateladze and the other anonymous reviewer(s) for their contribution to the peer review of this work.

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    This file contains Supplementary Figure 1, the uncropped scans with size marker indications.

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DOI

https://doi.org/10.1038/nature18312

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