Abstract
The DNA-damage response (DDR) arrests cell-cycle progression until damage is removed. DNA-damage-induced cellular senescence is associated with persistent DDR. The molecular bases that distinguish transient from persistent DDR are unknown. Here we show that a large fraction of exogenously induced persistent DDR markers is associated with telomeric DNA in cultured cells and mammalian tissues. In yeast, a chromosomal DNA double-strand break next to a telomeric sequence resists repair and impairs DNA ligase 4 recruitment. In mammalian cells, ectopic localization of telomeric factor TRF2 next to a double-strand break induces persistent DNA damage and DDR. Linear, but not circular, telomeric DNA or scrambled DNA induces a prolonged checkpoint in normal cells. In terminally differentiated tissues of old primates, DDR markers accumulate at telomeres that are not critically short. We propose that linear genomes are not uniformly reparable and that telomeric DNA tracts, if damaged, are irreparable and trigger persistent DDR and cellular senescence.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Buy this article
Purchase on Springer Link
Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
Accession codes
Change history
18 April 2012
In the version of this Article initially published online and in print, a reference was inadvertently omitted.
References
Campisi, J. & d’Adda di Fagagna, F. Cellular senescence: when bad things happen to good cells. Nat. Rev. Mol. Cell Biol. 8, 729–740 (2007).
Collado, M., Blasco, M. A. & Serrano, M. Cellular senescence in cancer and aging. Cell 130, 223–233 (2007).
Braig, M. et al. Oncogene-induced senescence as an initial barrier in lymphoma development. Nature 436, 660–665 (2005).
Chen, Z. et al. Crucial role of p53-dependent cellular senescence in suppression of Pten-deficient tumorigenesis. Nature 436, 725–730 (2005).
Collado, M. et al. Tumour biology: senescence in premalignant tumours. Nature 436, 642 (2005).
Michaloglou, C. et al. BRAFE600-associated senescence-like cell cycle arrest of human naevi. Nature 436, 720–724 (2005).
Baker, D. J. et al. Clearance of p16Ink4a-positive senescent cells delays ageing-associated disorders. Nature 479, 232–236 (2011).
d’Adda di Fagagna, F. et al. A DNA damage checkpoint response in telomere-initiated senescence. Nature 426, 194–198 (2003).
Herbig, U., Jobling, W. A., Chen, B. P., Chen, D. J. & Sedivy, J. M. Telomere shortening triggers senescence of human cells through a pathway involving ATM, p53, and p21(CIP1), but not p16(INK4a). Mol. Cell 14, 501–513 (2004).
Harley, C. B., Futcher, A. B. & Greider, C. W. Telomeres shorten during ageing of human fibroblasts. Nature 345, 458–460 (1990).
Evan, G. I. & d’Adda di Fagagna, F. Cellular senescence: hot or what? Curr. Opin. Genet. Dev. 19, 25–31 (2009).
Halazonetis, T. D., Gorgoulis, V. G. & Bartek, J. An oncogene-induced DNA damage model for cancer development. Science 319, 1352–1355 (2008).
Schmitt, C. A. Senescence, apoptosis and therapy-cutting the lifelines of cancer. Nat. Rev. Cancer 3, 286–295 (2003).
Herbig, U., Ferreira, M., Condel, L., Carey, D. & Sedivy, J. M. Cellular senescence in aging primates. Science 311, 1257 (2006).
Rossi, D. J. et al. Deficiencies in DNA damage repair limit the function of haematopoietic stem cells with age. Nature 447, 725–729 (2007).
Nijnik, A. et al. DNA repair is limiting for haematopoietic stem cells during ageing. Nature 447, 686–690 (2007).
Jeyapalan, J. C., Ferreira, M., Sedivy, J. M. & Herbig, U. Accumulation of senescent cells in mitotic tissue of aging primates. Mech. Ageing Dev. 128, 36–44 (2007).
Jackson, S. P. & Bartek, J. The DNA-damage response in human biology and disease. Nature 461, 1071–1078 (2009).
Meier, A. et al. Spreading of mammalian DNA-damage response factors studied by ChIP-chip at damaged telomeres. EMBO J. 26, 2707–2718 (2007).
Zhou, B. B. & Bartek, J. Targeting the checkpoint kinases: chemosensitization versus chemoprotection. Nat. Rev. Cancer 4, 216–225 (2004).
Rodier, F. et al. DNA-SCARS: distinct nuclear structures that sustain damage-induced senescence growth arrest and inflammatory cytokine secretion. J. Cell Sci. 124, 68–81 (2011).
O’Sullivan, R. J. & Karlseder, J. Telomeres: protecting chromosomes against genome instability. Nat. Rev. Mol. Cell Biol. 11, 171–181 (2010).
Bae, N. S. & Baumann, P. A RAP1/TRF2 complex inhibits nonhomologous end-joining at human telomeric DNA ends. Mol. Cell 26, 323–334 (2007).
Bombarde, O. et al. TRF2/RAP1 and DNA-PK mediate a double protection against joining at telomeric ends. EMBO J. 29, 1573–1584 (2010).
Hewitt, G. et al. Telomeres are favoured targets of a persistent DNA damage response in ageing and stress-induced senescence. Nat. Commun. 3, 708 (2012).
Le, O. N. et al. Ionizing radiation-induced long-term expression of senescence markers in mice is independent of p53 and immune status. Aging Cell 9, 398–409 (2010).
Van Steensel, B., Smogorzewska, A. & de Lange, T. TRF2 protects human telomeres from end-to-end fusions. Cell 92, 401–413 (1998).
Takai, H., Smogorzewska, A. & de Lange, T. DNA damage foci at dysfunctional telomeres. Curr. Biol. 13, 1549–1556 (2003).
Fujita, K. et al. Positive feedback between p53 and TRF2 during telomere-damage signalling and cellular senescence. Nat. Cell Biol. 12, 1205–1212 (2010).
Gonzalo, S. et al. Role of the RB1 family in stabilizing histone methylation at constitutive heterochromatin. Nat. Cell Biol. 7, 420–428 (2005).
Marchion, D. C., Bicaku, E., Daud, A. I., Sullivan, D. M. & Munster, P. N. Valproic acid alters chromatin structure by regulation of chromatin modulation proteins. Cancer Res. 65, 3815–3822 (2005).
Goodarzi, A. A. et al. ATM signaling facilitates repair of DNA double-strand breaks associated with heterochromatin. Mol. Cell 31, 167–177 (2008).
Diede, S. J. & Gottschling, D. E. Exonuclease activity is required for sequence addition and Cdc13p loading at a de novo telomere. Curr. Biol. 11, 1336–1340 (2001).
Michelson, R. J., Rosenstein, S. & Weinert, T. A telomeric repeat sequence adjacent to a DNA double-stranded break produces an anticheckpoint. Genes Dev. 19, 2546–2559 (2005).
Marcand, S., Pardo, B., Gratias, A., Cahun, S. & Callebaut, I. Multiple pathways inhibit NHEJ at tolemeres. Genes Dev. 22, 1153–1158 (2008).
Celli, G. B. & de Lange, T. DNA processing is not required for ATM-mediated telomere damage response after TRF2 deletion. Nat. Cell Biol. 7, 712–718 (2005).
Ancelin, K. et al. Targeting assay to study the cis functions of human telomeric proteins: evidence for inhibition of telomerase by TRF1 and for activation of telomere degradation by TRF2. Mol. Cell Biol. 22, 3474–3487 (2002).
Soutoglou, E. et al. Positional stability of single double-strand breaks in mammalian cells. Nat. Cell Biol. 9, 675–682 (2007).
Huang, L. C., Clarkin, K. C. & Wahl, G. M. Sensitivity and selectivity of the DNA damage sensor responsible for activating p53-dependent G1 arrest. Proc. Natl Acad. Sci. USA 93, 4827–4832 (1996).
Petersen, S., Saretzki, G. & von Zglinicki, T. Preferential accumulation of single-stranded regions in telomeres of human fibroblasts. Exp. Cell Res. 239, 152–160 (1998).
Rochette, P. J. & Brash, D. E. Human telomeres are hypersensitive to UV-induced DNA damage and refractory to repair. PLoS Genet. 6, e1000926 (2010).
Gomes, N. M. et al. Comparative biology of mammalian telomeres: hypotheses on ancestral states and the roles of telomeres in longevity determination. Aging Cell 10, 761–768 (2011).
Giaimo, S. & d’Adda di Fagagna, F. Is cellular senescence an example of antagonistic pleiotropy? Aging Celldoi: 10.1111/j.1474-9726.2012.00807.x (2012).
Marusyk, A., Wheeler, L. J., Mathews, C. K. & DeGregori, J. p53 mediates senescence-like arrest induced by chronic replicational stress. Mol. Cell Biol. 27, 5336–5351 (2007).
Di Micco, R. et al. Oncogene-induced senescence is a DNA damage response triggered by DNA hyper-replication. Nature 444, 638–642 (2006).
Ye, J. et al. TRF2 and apollo cooperate with topoisomerase 2α to protect human telomeres from replicative damage. Cell 142, 230–242 (2010).
Soutoglou, E. & Misteli, T. Activation of the cellular DNA damage response in the absence of DNA lesions. Science 320, 1507–1510 (2008).
Ziv, Y. et al. Chromatin relaxation in response to DNA double-strand breaks is modulated by a novel ATM- and KAP-1 dependent pathway. Nat. Cell Biol. 8, 870–876 (2006).
Francia, S., Weiss, R. S. & d’Adda di Fagagna, F. Need telomere maintenance? Call 911 Cell Div. 2, 3 (2007).
Ghisletti, S. et al. Identification and characterization of enhancers controlling the inflammatory gene expression program in macrophages. Immunity 32, 317–328 (2010).
Li, H. & Durbin, R. Fast and accurate long-read alignment with Burrows–Wheeler transform. Bioinformatics 26, 589–595 (2010).
Nobuyuki, O. A threshold selection method from gray-level histograms. IEEE Trans. Sys. Man. Cyber. 9, 62–66 (1979).
Duffy, D. L. Lodplot: plot a genome scan. R package version 1.1. (2007).
The R Development Core Team, R: A Language and Environment for Statistical Computing (R Foundation for Statistical Computing, Vienna, Austria, 2008.
Di Micco, R. et al. Interplay between oncogene-induced DNA damage response and heterochromatin in senescence and cancer. Nat. Cell Biol. 13, 292–302 (2011).
Viscardi, V., Bonetti, D., Cartagena-Lirola, H., Lucchini, G. & Longhese, M. P. MRX-dependent DNA damage response to short telomeres. Mol. Biol. Cell 18, 3047–3058 (2007).
Acknowledgements
We thank V. Dall’Olio and L. Tizzoni from IFOM RT–PCR Unit, A. Oldani and all the IFOM Imaging Unit, L. Rotta from IFOM Microarray and NGS Unit, IFOM Cell Biology Unit for support; V. Boccardi for discussions; P. Baumann, T. F. Halazonetis, T. Weinert, D. E. Gottschling, E. Soutoglou, E. Gilson, P. Jeggo, A. Musacchio and S. Minucci for sharing reagents; O. Le for mouse brain tissue sectioning and all F.d’A.d.F. laboratory members for discussions. F.d’A.d.F.’s laboratory is supported by FIRC (Fondazione Italiana per la Ricerca sul Cancro), AIRC (Associazione Italiana per la Ricerca sul Cancro; grant number 8866), European Union (GENINCA, contract number 202230), HFSP (Human Frontier Science Program), AICR (Association for International Cancer Research), EMBO Young Investigator Program and Telethon. M.P.L.’s laboratory is supported by AIRC (grant number 11407), Cofinanziamento 2008 MIUR/Università di Milano-Bicocca and the European Union. C.M.B. is supported by a grant from the Canadian Institute of Health Research (number IAO-79317). U.H. is supported by a New Scholar Award from the Ellison Medical Foundation (AG-NS-0387-07) and by a grant (R01CA136533) from the National Cancer Institute.
Author information
Authors and Affiliations
Contributions
F.R. generated and assembled data in Figs 5a–c,e–f,h–i, 7a–c, 8d and Supplementary Figs S1b–c, S3b, S4c, S5, S6, S7b–c, S8a; M.C. and M.P.L. generated data in Fig. 6; S.B. carried out the microinjection experiments; D.C. carried out the analysis of sequencing data and generated data in Fig. 3a,b; J.M.K. generated data in Fig. 8e and Supplementary Fig. S8b; G.B. contributed to the pre-processing and analysis of sequencing data in Fig. 3a,b; M.D. provided technical assistance; V.M. generated data in Fig. 5d,g and provided technical assistance; C.M.B. provided irradiated mouse brain sections; U.H. provided baboon sections and edited the manuscript; M.F. generated and assembled data of all remaining figures, carried out ChIP assays in mammalian cells and contributed to experimental design and manuscript writing; F.d’A.d.F. planned and supervised the project and wrote the manuscript.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary Information
Supplementary Information (PDF 3555 kb)
Rights and permissions
About this article
Cite this article
Fumagalli, M., Rossiello, F., Clerici, M. et al. Telomeric DNA damage is irreparable and causes persistent DNA-damage-response activation. Nat Cell Biol 14, 355–365 (2012). https://doi.org/10.1038/ncb2466
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/ncb2466
This article is cited by
-
Senescence-related impairment of autophagy induces toxic intraneuronal amyloid-β accumulation in a mouse model of amyloid pathology
Acta Neuropathologica Communications (2023)
-
Sex- and age-related differences in renal and cardiac injury and senescence in stroke-prone spontaneously hypertensive rats
Biology of Sex Differences (2023)
-
Regulation of self-renewal and senescence in primitive mesenchymal stem cells by Wnt and TGFβ signaling
Stem Cell Research & Therapy (2023)
-
Senescence atlas reveals an aged-like inflamed niche that blunts muscle regeneration
Nature (2023)
-
Longitudinal telomere dynamics within natural lifespans of a wild bird
Scientific Reports (2023)