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Tel1ATM and Rad3ATR kinases promote Ccq1-Est1 interaction to maintain telomeres in fission yeast

Nature Structural & Molecular Biology volume 18, pages 1408–1413 (2011)Cite this article

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Abstract

The evolutionarily conserved shelterin complex has been shown to play both positive and negative roles in telomerase regulation in mammals and fission yeast. Although shelterin prevents the checkpoint kinases ATM and ATR from fully activating DNA damage responses at telomeres in mammalian cells, those kinases also promote telomere maintenance. In fission yeast, cells lacking both Tel1 (ATM ortholog) and Rad3 (ATR ortholog) fail to recruit telomerase to telomeres and survive by circularizing chromosomes. However, the critical telomere substrate(s) of Tel1ATM and Rad3ATR was unknown. Here we show that phosphorylation of the shelterin subunit Ccq1 on Thr93, redundantly mediated by Tel1ATM and/or Rad3ATR, is essential for telomerase association with telomeres. In addition, we show that the telomerase subunit Est1 interacts directly with the phosphorylated Thr93 of Ccq1 to ensure telomere maintenance. The shelterin subunits Taz1, Rap1 and Poz1 (previously established inhibitors of telomerase) were also found to negatively regulate Ccq1 phosphorylation. These findings establish Tel1ATM/Rad3ATR-dependent Ccq1 Thr93 phosphorylation as a critical regulator of telomere maintenance in fission yeast.

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Figure 1: Ccq1 interacts with both Tpz1 and Est1.
Figure 2: The phosphopeptide-binding motif of Est1 is important for telomere maintenance.
Figure 3: Thr93 of Ccq1 is essential for telomere maintenance.
Figure 4: Ccq1-Est1 interaction is dependent on phosphorylated Ccq1 Thr93.

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Acknowledgements

We thank F. Ishikawa (Kyoto University), J.P. Cooper (Cancer Research UK), M.R. Flory (Mendel Biotechnology, Inc.), V.A. Zakian (Princeton University), A.M. Carr (University of Sussex) and P. Russell (The Scripps Research Institute) for sharing published strains and plasmids, L. Khair for her initial efforts to generate yeast strains and reagents used in this study, and P. Baumann (Stowers Institute) for generously sharing his unpublished anti-Ccq1 antibody. We also thank F. Ishikawa for communicating unpublished results. J.K. was supported in part by the Federal Work-Study program. This work was supported by US National Institutes of Health grant GM078253 (T.M.N.).

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  1. Department of Biochemistry and Molecular Genetics, College of Medicine, University of Illinois at Chicago, Chicago, Illinois, USA

    Bettina A Moser, Ya-Ting Chang, Jorgena Kosti & Toru M Nakamura

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  1. Bettina A Moser
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Contributions

B.A.M. designed, performed and analyzed most of the experiments in this study and wrote the paper. Y.-T.C. performed the ChIP experiments illustrated in Figure 3a and initially observed Ccq1 hyperphosphorylation. J.K. assisted B.A.M. in construction of various yeast two-hybrid plasmids. T.M.N. conceived the study, designed and performed experiments, analyzed data and wrote the paper.

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Correspondence to Toru M Nakamura.

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Moser, B., Chang, YT., Kosti, J. et al. Tel1ATM and Rad3ATR kinases promote Ccq1-Est1 interaction to maintain telomeres in fission yeast. Nat Struct Mol Biol 18, 1408–1413 (2011). https://doi.org/10.1038/nsmb.2187

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