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DNA processing is not required for ATM-mediated telomere damage response after TRF2 deletion

Abstract

Telomere attrition and other forms of telomere damage can activate the ATM kinase pathway. What generates the DNA damage signal at mammalian chromosome ends or at other double-strand breaks is not known. Telomere dysfunction is often accompanied by disappearance of the 3′ telomeric overhang1,2, raising the possibility that DNA degradation could generate the structure that signals2. Here we address these issues by studying telomere structure after conditional deletion of mouse TRF2, the protective factor at telomeres. Upon removal of TRF2 from TRF2F/− p53−/− mouse embryo fibroblasts, a telomere damage response is observed at most chromosome ends. As expected, the telomeres lose the 3′ overhang and are processed by the non-homologous end-joining pathway. Non-homologous end joining of telomeres was abrogated in DNA ligase IV-deficient (Lig4−/−) cells. Unexpectedly, the telomeres of TRF2−/− Lig4−/− p53−/− cells persisted in a free state without undergoing detectable DNA degradation. Notably, the telomeres retained their 3′ overhangs, but they were recognized as sites of DNA damage, accumulating the DNA damage response factors 53BP1 and γ-H2AX, and activating the ATM kinase. Thus, activation of the ATM kinase pathway at chromosome ends does not require overhang degradation or other overt DNA processing.

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Figure 1: Conditional deletion of mouse TRF2.
Figure 2: DNA ligase IV-mediated genome-wide telomere fusions upon TRF2 deletion.
Figure 3: Lack of degradation of telomeric DNA in TRF2−/− Lig4−/− cells.
Figure 4: DNA damage response upon TRF2 deletion from TRF2F/− Lig4+/+ p53−/− and TRF2F/− Lig4−/− p53−/− cells.

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Acknowledgements

We gratefully acknowledge D. White for mouse husbandry and the RU Gene Targeting and Transgenic facilities for help in generating genetically modified mouse strains. F. Alt and C. Canyon are thanked for providing Lig4−/− mice, and T. Jacks for p53−/− mice. T. Halazonetis, J. Petrini, Y. Shiloh, T. Jacks, D. Livingston and G. Hannon are thanked for providing antibodies and plasmids. This work was supported by an NIH grant (GM49046-12) to T.d.L., and by funding from The Rockefeller University's Women & Science Fellowship Program, and by The Leukemia & Lymphoma Society to G.B.C.

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Correspondence to Titia de Lange.

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Supplementary figures S1 - S3 and supplementary tables S1 and S2 (PDF 408 kb)

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Celli, G., de Lange, T. DNA processing is not required for ATM-mediated telomere damage response after TRF2 deletion. Nat Cell Biol 7, 712–718 (2005). https://doi.org/10.1038/ncb1275

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