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DNA-damage response and repair activities at uncapped telomeres depend on RNF8

Nature Cell Biology volume 13, pages 1139–1145 (2011)Cite this article

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Abstract

Loss of telomere protection causes natural chromosome ends to become recognized by DNA-damage response and repair proteins. These events result in ligation of chromosome ends with dysfunctional telomeres, thereby causing chromosomal aberrations on cell division. The control of these potentially dangerous events at deprotected chromosome ends with their unique telomeric chromatin configuration is poorly understood. In particular, it is unknown to what extent bulky modification of telomeric chromatin is involved. Here we show that uncapped telomeres accumulate ubiquitylated histone H2A in a manner dependent on the E3 ligase RNF8. The ability of RNF8 to ubiquitylate telomeric chromatin is associated with its capacity to facilitate accumulation of both 53BP1 and phospho-ATM at uncapped telomeres and to promote non-homologous end-joining of deprotected chromosome ends. In line with the detrimental effect of RNF8 on uncapped telomeres, depletion of RNF8, as well as of the E3 ligase RNF168, reduces telomere-induced genome instability. This indicates that, besides suppressing tumorigenesis by mediating repair of DNA double-strand breaks, RNF8 and RNF168 might enhance cancer development by aggravating telomere-induced genome instability.

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Figure 1: RNF8 contributes to telomere-induced genome instability.
Figure 2: RNF8 facilitates telomeric G-overhang degradation and telomere fusion.
Figure 3: Impaired 53BP1 and p-ATM accumulation at uncapped telomeres on knockdown of Rnf8.
Figure 4: Telomere uncapping induces ubiquitylation of H2A and H2AX in an RNF8-dependent manner.
Figure 5: 53BP1, p-ATM and Ub–H2A accumulation at uncapped telomeres and telomere-induced genome instability depend on the RNF8 FHA and E3-ligase domains.

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Acknowledgements

We thank A. Konishi, G. Celli and T. de Lange for the Trf2ts allele, Trf2flox/−; p53−/− MEFs and Trf2flox/−; p53−/−; Lig4−/− MEFs, J. van der Torre and K. Beekhuis for help with generating and testing Trf2−/−; p53−/−;TRF2ts clones, J. van Noord for technical assistance at the beginning of this project, S. Burkhard for help with RNF168 qRT–PCR, D. Silver and D. Livingston for the HR-Cre plasmid, F. Mattiroli for pCDNA-HA-RNF8, S. Kabir (de Lange laboratory) for technical advice on IF-FISH and FISH and L. Oomen and L. Brocks of the NKI digital microscope facility for help with confocal microscope imaging. We also thank J. van der Torre and S. Burkhard for sharing unpublished results on the identification of RNF8 shRNAs in a genome-wide shRNA library screen in TRF2ts cells and E. Citterio and A. Berns for critically reading this manuscript. This work was supported by grants NKI 2005-3458 and NKI 2007-3907 from the Dutch Cancer Society to J.J.L.J.

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  1. Division of Molecular Genetics, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands

    Marieke H. Peuscher & Jacqueline J. L. Jacobs

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  1. Marieke H. Peuscher
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Contributions

M.H.P. and J.J.L.J. designed the experiments. M.H.P. carried out most experimental work. J.J.L.J. supervised the project, helped generate Trf2−/−; p53−/−;TRF2ts clones and carried out all molecular cloning involved and experiments represented in Fig. 3d and Supplementary Fig. S4. J.J.L.J. wrote the manuscript.

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Correspondence to Jacqueline J. L. Jacobs.

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Peuscher, M., Jacobs, J. DNA-damage response and repair activities at uncapped telomeres depend on RNF8. Nat Cell Biol 13, 1139–1145 (2011). https://doi.org/10.1038/ncb2326

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