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Anticheckpoint pathways at telomeres in yeast

Nature Structural & Molecular Biology volume 19, pages 307–313 (2012)Cite this article

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Abstract

Telomeres hide (or 'cap') chromosome ends from DNA-damage surveillance mechanisms that arrest the cell cycle and promote repair, but the checkpoint status of telomeres is not well understood. Here we characterize the response in Saccharomyces cerevisiae to DNA double-strand breaks (DSBs) flanked by varying amounts of telomeric repeat sequences (TG1–3). We show that even short arrays of TG1–3 repeats do not induce G2/M arrest. Both Rif1 and Rif2 are required for capping at short, rapidly elongating ends, yet are largely dispensable for protection of longer telomeric arrays. Rif1 and Rif2 act through parallel pathways to block accumulation of both RPA and Rad24, activators of checkpoint kinase Mec1 (ATR). Finally, we show that Rif function is correlated with an 'anticheckpoint' effect, in which checkpoint recovery at an adjacent unprotected end is stimulated, and we provide insight into the molecular mechanism of this phenomenon.

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Figure 1: Suppression of G2/M arrest at a DSB flanked by short (80 bp) telomeric repeats is RIF1 and RIF2 dependent.
Figure 2: Inactivation of RIF1 or RIF2 increases the recruitment of proteins involved in the DNA damage response.
Figure 3: Telomeric tracts elicit an anticheckpoint.
Figure 4: The presence of telomeric tracts on the distal part of a break influences the recruitment of proteins on the proximal part.

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Acknowledgements

We thank C. Follonier for initiating the project, P. Damay for help in plasmid construction, B. Knight for helpful discussions concerning statistical analysis, S. Brill (Rutgers, State University of New Jersey) for Rfa2-specific antibody, A. Pellicioli (University of Milan) for Rad53-specific antibody, N. Roggli for artwork, V. Ribaud (Shore laboratory) for strains (listed in Supplementary Table 1), members of the Shore laboratory for discussions and suggestions, and A. Bianchi for discussions and critical reading of the manuscript. These studies were supported by a grant from the Swiss National Fund to D.S. (31003A-116716), by the National Center for Competence in Research 'Frontiers in Genetics' program of the Swiss National Fund, and by the Canton and Republic of Geneva. C.R. was supported by a postdoctoral fellowship from the Fondation pour la Recherche Médicale (France).

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  1. Cyril Ribeyre

    Present address: Present address: Institute of Human Genetics, Centre National de la Recherche Scientifique UPR1142, Montpellier, France.,

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  1. Department of Molecular Biology and National Center for Competence in Research Program 'Frontiers in Genetics', University of Geneva, Geneva, Switzerland

    Cyril Ribeyre & David Shore

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C.R. and D.S. designed the experiments and analyzed the data. C.R. performed the experiments. C.R. and D.S. wrote the manuscript.

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Correspondence to David Shore.

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Ribeyre, C., Shore, D. Anticheckpoint pathways at telomeres in yeast. Nat Struct Mol Biol 19, 307–313 (2012). https://doi.org/10.1038/nsmb.2225

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