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Chfr and RNF8 synergistically regulate ATM activation

Nature Structural & Molecular Biology volume 18, pages 761–768 (2011)Cite this article

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Abstract

Protein ubiquitination is a crucial component of the DNA damage response. To study the mechanism of the DNA damage–induced ubiquitination pathway, we analyzed the impact of the loss of two E3 ubiquitin ligases, RNF8 and Chfr. Notably, DNA damage–induced activation of ATM kinase is suppressed in cells deficient in both RNF8 and Chfr (double-knockout, or DKO), and DKO mice develop thymic lymphomas that are nearly diploid but harbor clonal chromosome translocations. Moreover, DKO mice and cells are hypersensitive to ionizing radiation. We present evidence that RNF8 and Chfr synergistically regulate histone ubiquitination to control histone H4 Lys16 acetylation through MRG15-dependent acetyltransferase complexes. Through these complexes, RNF8 and Chfr affect chromatin relaxation and modulate ATM activation and DNA damage response pathways. Collectively, our findings demonstrate that two chromatin-remodeling factors, RNF8 and Chfr, function together to activate ATM and maintain genomic stability in vivo.

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Figure 1: The DNA damage–induced ATM signaling pathway is impaired in DKO cells.
Figure 2: RNF8 and Chfr double-deficient mice develop T-cell lymphoma.
Figure 3: Thymic lymphomas from DKO mice harbor clonal translocations.
Figure 4: DKO mice and MEFs are hypersensitive to ionizing radiation.
Figure 5: RNF8 and Chfr synergistically regulate histone ubiquitination and acetylation.
Figure 6: MRG15 links histone H2B ubiquitination and H4K16 acetylation.
Figure 7: Suppression of histone acetylation rescues ATM-dependent DNA damage response in DKO MEFs.
Figure 8: A model of histone modifications, chromatin relaxation and ATM activation after DNA damage.

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Acknowledgements

We thank E. Fearon, K. Cho, B. Margolis, D. Yali and L. Yang at the University of Michigan for sharing experimental equipment, Z. You at Washington University and K. Tominaga at University of Texas Health Science Center at San Antonio for reagents and J. Keller for proofreading the manuscript. This work was supported by the American Cancer Society (RSG-08-125-01-CCG to X.Y.) and the US National Institutes of Health (CA132755 and CA130899 to X.Y.). X.Y. is a recipient of the Era of Hope Scholar Award from the US Department of Defense.

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Authors and Affiliations

  1. Division of Molecular Medicine and Genetics, Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, Michigan, USA

    Jiaxue Wu, Yibin Chen, Lin-Yu Lu & Xiaochun Yu

  2. Department of Pathology, University of Michigan Medical School, Ann Arbor, Michigan, USA

    Yipin Wu & David O Ferguson

  3. Department of Radiation Oncology, University of Michigan Medical School, Ann Arbor, Michigan, USA

    Michelle T Paulsen & Mats Ljungman

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Contributions

J.W. performed most experiments. Y.C. analyzed the MRG15-related protein-protein interactions. L.-Y.L., M.T.P., M.L., Y.W. and D.O.F. provided technical support for various assays. X.Y. designed the experiments. X.Y. and J.W. wrote the manuscript. All the authors read and approved the final manuscript.

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Correspondence to Xiaochun Yu.

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Wu, J., Chen, Y., Lu, LY. et al. Chfr and RNF8 synergistically regulate ATM activation. Nat Struct Mol Biol 18, 761–768 (2011). https://doi.org/10.1038/nsmb.2078

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