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Telomeres avoid end detection by severing the checkpoint signal transduction pathway

Nature volume 467, pages 228–232 (2010)Cite this article

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Abstract

Telomeres protect the normal ends of chromosomes from being recognized as deleterious DNA double-strand breaks. Recent studies have uncovered an apparent paradox: although DNA repair is prevented, several proteins involved in DNA damage processing and checkpoint responses are recruited to telomeres in every cell cycle and are required for end protection1. It is currently not understood how telomeres prevent DNA damage responses from causing permanent cell cycle arrest. Here we show that fission yeast (Schizosaccharomyces pombe) cells lacking Taz1, an orthologue of human TRF1 and TRF2 (ref. 2), recruit DNA repair proteins (Rad22RAD52 and Rhp51RAD51, where the superscript indicates the human orthologue) and checkpoint sensors (RPA, Rad9, Rad26ATRIP and Cut5/Rad4TOPBP1) to telomeres. Despite this, telomeres fail to accumulate the checkpoint mediator Crb253BP1 and, consequently, do not activate Chk1-dependent cell cycle arrest. Artificially recruiting Crb253BP1 to taz1Δ telomeres results in a full checkpoint response and cell cycle arrest. Stable association of Crb253BP1 to DNA double-strand breaks requires two independent histone modifications: H4 dimethylation at lysine 20 (H4K20me2) and H2A carboxy-terminal phosphorylation (γH2A)3,4,5. Whereas γH2A can be readily detected, telomeres lack H4K20me2, in contrast to internal chromosome locations. Blocking checkpoint signal transduction at telomeres requires Pot1 and Ccq1, and loss of either Pot1 or Ccq1 from telomeres leads to Crb253BP1 foci formation, Chk1 activation and cell cycle arrest. Thus, telomeres constitute a chromatin-privileged region of the chromosomes that lack essential epigenetic markers for DNA damage response amplification and cell cycle arrest. Because the protein kinases ATM and ATR must associate with telomeres in each S phase to recruit telomerase6, exclusion of Crb253BP1 has a critical role in preventing telomeres from triggering cell cycle arrest.

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Figure 1: taz1 Δ telomeres undergo the DNA damage response without eliciting a checkpoint-dependent cell cycle arrest.
Figure 2: taz1 Δ telomeres initiate a DNA damage checkpoint response.
Figure 3: Dysfunctional taz1 Δ telomeres avert cell cycle arrest by preventing recruitment of Crb253BP1.
Figure 4: Pot1 and Ccq1 prevent Crb2 53BP1 -dependent checkpoints at telomeres.

References

  1. de Lange, T. How telomeres solve the end-protection problem. Science 326, 948–952 (2009)

    Article  ADS  CAS  Google Scholar 

  2. Li, B., Oestreich, S. & de Lange, T. Identification of human Rap1: implications for telomere evolution. Cell 101, 471–483 (2000)

    Article  CAS  Google Scholar 

  3. Du, L.-L., Nakamura, T. M. & Russell, P. Histone modification-dependent and -independent pathways for recruitment of checkpoint protein Crb2 to double-strand breaks. Genes Dev. 20, 1583–1596 (2006)

    Article  CAS  Google Scholar 

  4. Greeson, N. T., Sengupta, R., Arida, A. R., Jenuwein, T. & Sanders, S. L. Di-methyl H4 lysine 20 targets the checkpoint protein Crb2 to sites of DNA damage. J. Biol. Chem. 283, 33168–33174 (2008)

    Article  CAS  Google Scholar 

  5. Nakamura, T. M., Du, L.-L., Redon, C. & Russell, P. Histone H2A phosphorylation controls Crb2 recruitment at DNA breaks, maintains checkpoint arrest, and influences DNA repair in fission yeast. Mol. Cell. Biol. 24, 6215–6230 (2004)

    Article  CAS  Google Scholar 

  6. Moser, B. A., Subramanian, L., Khair, L., Chang, Y.-T. & Nakamura, T. M. Fission yeast Tel1ATM and Rad3ATR promote telomere protection and telomerase recruitment. PLoS Genet. 5, e1000622 (2009)

    Article  Google Scholar 

  7. Ferreira, M. G. & Cooper, J. P. The fission yeast Taz1 protein protects chromosomes from Ku-dependent end-to-end fusions. Mol. Cell 7, 55–63 (2001)

    Article  CAS  Google Scholar 

  8. Ferreira, M. G. & Cooper, J. P. Two modes of DNA double-strand break repair are reciprocally regulated through the fission yeast cell cycle. Genes Dev. 18, 2249–2254 (2004)

    Article  CAS  Google Scholar 

  9. Miller, K. M., Rog, O. & Cooper, J. P. Semi-conservative DNA replication through telomeres requires Taz1. Nature 440, 824–828 (2006)

    Article  ADS  CAS  Google Scholar 

  10. Rog, O., Miller, K. M., Ferreira, M. G. & Cooper, J. P. Sumoylation of RecQ helicase controls the fate of dysfunctional telomeres. Mol. Cell 33, 559–569 (2009)

    Article  CAS  Google Scholar 

  11. Tomita, K. et al. Fission yeast Dna2 is required for generation of the telomeric single-strand overhang. Mol. Cell. Biol. 24, 9557–9567 (2004)

    Article  CAS  Google Scholar 

  12. Churikov, D. & Price, C. M. Pot1 and cell cycle progression cooperate in telomere length regulation. Nature Struct. Mol. Biol. 15, 79–84 (2007)

    Article  Google Scholar 

  13. Denchi, E. L. & de Lange, T. Protection of telomeres through independent control of ATM and ATR by TRF2 and POT1. Nature 448, 1068–1071 (2007)

    Article  ADS  CAS  Google Scholar 

  14. Guo, X. et al. Dysfunctional telomeres activate an ATM-ATR-dependent DNA damage response to suppress tumorigenesis. EMBO J. 26, 4709–4719 (2007)

    Article  CAS  Google Scholar 

  15. Moser, B. A. et al. Differential arrival of leading and lagging strand DNA polymerases at fission yeast telomeres. EMBO J. 28, 810–820 (2009)

    Article  CAS  Google Scholar 

  16. Verdun, R. E., Crabbe, L., Haggblom, C. & Karlseder, J. Functional human telomeres are recognized as DNA damage in G2 of the cell cycle. Mol. Cell 20, 551–561 (2005)

    Article  CAS  Google Scholar 

  17. Mochida, S. et al. Regulation of checkpoint kinases through dynamic interaction with Crb2. EMBO J. 23, 418–428 (2004)

    Article  CAS  Google Scholar 

  18. Sanders, S. L. et al. Methylation of histone H4 lysine 20 controls recruitment of Crb2 to sites of DNA damage. Cell 119, 603–614 (2004)

    Article  CAS  Google Scholar 

  19. Wang, Y. & Jia, S. Degrees make all the difference: the multifunctionality of histone H4 lysine 20 methylation. Epigenetics 4, 273–276 (2009)

    Article  CAS  Google Scholar 

  20. Chikashige, Y. & Hiraoka, Y. Telomere binding of the Rap1 protein is required for meiosis in fission yeast. Curr. Biol. 11, 1618–1623 (2001)

    Article  CAS  Google Scholar 

  21. Miller, K. M., Ferreira, M. G. & Cooper, J. P. Taz1, Rap1 and Rif1 act both interdependently and independently to maintain telomeres. EMBO J. 24, 3128–3135 (2005)

    Article  CAS  Google Scholar 

  22. Tomita, K. & Cooper, J. P. Fission yeast Ccq1 is telomerase recruiter and local checkpoint controller. Genes Dev. 22, 3461–3474 (2008)

    Article  CAS  Google Scholar 

  23. Miyoshi, T., Kanoh, J., Saito, M. & Ishikawa, F. Fission yeast Pot1-Tpp1 protects telomeres and regulates telomere length. Science 320, 1341–1344 (2008)

    Article  ADS  CAS  Google Scholar 

  24. 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)

    Article  CAS  Google Scholar 

  25. Abdallah, P. et al. A two-step model for senescence triggered by a single critically short telomere. Nature Cell Biol. 11, 988–993 (2009)

    Article  CAS  Google Scholar 

  26. Hirano, Y. & Sugimoto, K. Cdc13 telomere capping decreases Mec1 association but does not affect Tel1 association with DNA ends. Mol. Biol. Cell 18, 2026–2036 (2007)

    Article  CAS  Google Scholar 

  27. Negrini, S., Ribaud, V., Bianchi, A. & Shore, D. DNA breaks are masked by multiple Rap1 binding in yeast: implications for telomere capping and telomerase regulation. Genes Dev. 21, 292–302 (2007)

    Article  CAS  Google Scholar 

  28. Botuyan, M. V. et al. Structural basis for the methylation state-specific recognition of histone H4-K20 by 53BP1 and Crb2 in DNA repair. Cell 127, 1361–1373 (2006)

    Article  CAS  Google Scholar 

  29. Goudsouzian, L. K., Tuzon, C. T. & Zakian, V. A. S. cerevisiae Tel1p and Mre11p are required for normal levels of Est1p and Est2p telomere association. Mol. Cell 24, 603–610 (2006)

    Article  CAS  Google Scholar 

  30. Verdun, R. E. & Karlseder, J. The DNA damage machinery and homologous recombination pathway act consecutively to protect human telomeres. Cell 127, 709–720 (2006)

    Article  CAS  Google Scholar 

  31. Bähler, J. et al. Heterologous modules for efficient and versatile PCR-based gene targeting in Schizosaccharomyces pombe . Yeast 14, 943–951 (1998)

    Article  Google Scholar 

  32. Moreno, S., Klar, A. & Nurse, P. Molecular genetic analysis of fission yeast Schizosaccharomyces pombe . Methods Enzymol. 194, 795–823 (1991)

    Article  CAS  Google Scholar 

  33. Khair, L., Subramanian, L., Moser, B. A. & Nakamura, T. M. Roles of heterochromatin and telomere proteins in regulation of fission yeast telomere recombination and telomerase recruitment. J. Biol. Chem. 285, 5327–5337 (2010)

    Article  CAS  Google Scholar 

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Acknowledgements

We thank J. Cooper, K. Tomita and the rest of the Telomere Biology Laboratory (Cancer Research UK, London) for support at the start of this project. We thank J. Cooper, T. Wolkow, P. Russell and A. M. Carr for strains and plasmids. We thank W. Kaufman for her initial effort in the establishment of HO strains. We are grateful to S. Grewal for sharing unpublished results and to D. Lydall for insights on the quantitative analysis of checkpoint activation. We thank K. Labib, L. Jansen, K. Xavier, R. Martinho and S. Lopes for critically reading the manuscript. T.C. and C.C.R. are supported by Fundação para a Ciência e a Tecnologia (FCT) postdoctoral fellowships. T.M.N. was supported by the Sidney Kimmel Scholar Program and his laboratory is supported by NIH grant GM078253. This work was supported by the FCT (PTDC/BIA-BCM/67261/2006) and the Association for International Cancer Research (06-396).

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Authors and Affiliations

  1. Instituto Gulbenkian de Ciência, Oeiras 2781-901, Portugal

    Tiago Carneiro, Clara C. Reis, Vanessa Borges & Miguel Godinho Ferreira

  2. Department of Biochemistry and Molecular Genetics, University of Illinois, Chicago, 60607, Illinois, USA

    Lyne Khair, Bettina A. Moser & Toru M. Nakamura

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  1. Tiago Carneiro
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  7. Miguel Godinho Ferreira
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Contributions

T.C. helped with the design and executed most experiments. L.K. performed ChIP experiments. C.C.R. performed western and Southern blotting experiments. V.B. performed live cell analysis. B.A.M. established the ChIP and HO assays. T.M.N. contributed to the design of the ChIP and HO assays. All authors contributed with strains and data analysis. M.G.F. conceived the study, performed live cell analysis and wrote the paper.

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Correspondence to Miguel Godinho Ferreira.

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Carneiro, T., Khair, L., Reis, C. et al. Telomeres avoid end detection by severing the checkpoint signal transduction pathway. Nature 467, 228–232 (2010). https://doi.org/10.1038/nature09353

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