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The DNA damage response at eroded telomeres and tethering to the nuclear pore complex

Nature Cell Biology volume 11, pages 980–987 (2009)Cite this article

Abstract

The ends of linear eukaryotic chromosomes are protected by telomeres, which serve to ensure proper chromosome replication and to prevent spurious recombination at chromosome ends. In this study, we show by single cell analysis that in the absence of telomerase, a single short telomere is sufficient to induce the recruitment of checkpoint and recombination proteins. Notably, a DNA damage response at eroded telomeres starts many generations before senescence and is characterized by the recruitment of Cdc13 (cell division cycle 13), replication protein A, DNA damage checkpoint proteins and the DNA repair protein Rad52 into a single focus. Moreover, we show that eroded telomeres, although remaining at the nuclear periphery, move to the nuclear pore complex. Our results link the DNA damage response at eroded telomeres to changes in subnuclear localization and suggest the existence of collapsed replication forks at eroded telomeres.

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Figure 1: Localization of Cdc13 and Rad52 to short telomeres.
Figure 2: Cdc13–Rad52 focus formation correlates with short telomeres and single-stranded DNA formation.
Figure 3: Recruitment of Cdc13, checkpoint and DNA-repair proteins to telomeres in senescing cells.
Figure 4: Eroded telomeres relocalize to the nuclear pore complex.
Figure 5: Tethering of telomeres to the NPC in senescing cells.

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Acknowledgements

We thank members of the Géli, Gilson and Lisby laboratories for helpful discussions concerning this work, A. Nicolas, X. Zhao, R. Rothstein, B. Palancade, V. Lundblad and R. Wellinger for sharing reagents and for fruitful discussions, S. Larose and R. Wellinger, who engineered the pTet-off-TLC1 construct, and S. Brill for the anti-RPA antibody. This work was supported by The Danish Agency for Science, Technology and Innovation (M.L.), the Villum Kann Rasmussen Foundation (M.L.), the Deutscher Akademischer Austausch Dienst (S.M.G.) and the Lundbeck Foundation (N.E.B.). The Agence Nationale de la Recherche (ANR programme blanc) and the Ligue Nationale Contre le Cancer (LNCC, équipes labelisées) supported V.G. and E.G. laboratories; B.K. is a recipient of a fellowship from the LNNC and P.A. is supported by the Lebanese National council for Scientific Research (CNRSL) and the Association pour la Recherche sur le Cancer (ARC).

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Author notes

  1. Basheer Khadaroo and M. Teresa Teixeira: These authors contributed equally to this work.

Authors and Affiliations

  1. CNRS, Unité Propre de Recherche 3081, Genome Instability and Carcinogenesis Conventionné par l'Université d'Aix-Marseille 2, 13402, Cedex 20, Marseille, France

    Basheer Khadaroo, Pierre Luciano, Marie Noelle Simon & Vincent Géli

  2. Laboratoire de Biologie Moléculaire de la Cellule, UMR 5239, CNRS/ ENS Lyon, Université Lyon 1, Ecole Normale Supérieure, 46 allée d'Italie, Cedex 07, F-69364 Lyon, France

    M. Teresa Teixeira, Pauline Abdallah & Eric Gilson

  3. Department of Biology, University of Copenhagen, DK-2200, Copenhagen N, Denmark

    Nadine Eckert-Boulet, Susanne M. Germann, Irene Gallina & Michael Lisby

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  1. Basheer Khadaroo
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Contributions

B.K., P.L. and M.N.S. performed the ChIP and telomeric blot analyses. N.E.B., I.G. and M.L. performed the microscopy. B.K., N.E.B., S.M.G., M.T.T. and P.A. constructed strains and assisted with data analysis. M.L. and P.L. performed telomere length PCR. V.G., M.L., E.G. and N.E.B. conceived and designed research, and wrote the manuscript.

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Correspondence to Vincent Géli.

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Khadaroo, B., Teixeira, M., Luciano, P. et al. The DNA damage response at eroded telomeres and tethering to the nuclear pore complex. Nat Cell Biol 11, 980–987 (2009). https://doi.org/10.1038/ncb1910

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