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Chromosome end protection plasticity revealed by Stn1p and Ten1p bypass of Cdc13p

Nature Cell Biology volume 8, pages 748–755 (2006)Cite this article

Abstract

Genome stability necessitates a mechanism to protect the termini of linear chromosomes from inappropriate degradation or recombination1,2. In many species this protection depends on 'capping' proteins that bind telomeric DNA3,4. The budding yeast Cdc13p binds single-stranded telomeric sequences5,6, prevents lethal degradation of chromosome ends7,8 and regulates telomere extension by telomerase5,9. Two Cdc13-interacting proteins, Stn1p and Ten1p, are also required for viability and telomere length regulation10,11. It has been proposed that Cdc13p DNA binding directs a Cdc13p–Stn1p–Ten1p complex to telomeres to mediate end protection12. However, the functional significance of these protein interactions, and their respective roles in maintaining telomere integrity, remain undefined. Here, we show that co-overexpressing TEN1 with a truncated form of STN1 efficiently bypasses the essential role of CDC13. We further show that this truncated Stn1p binds directly to Pol12p, a polymerase α-primase regulatory subunit, and that Pol12 activity is required for CDC13 bypass. Thus, Stn1p and Ten1p control a Cdc13p-independent telomere capping mechanism that is coupled to the conventional DNA replication machinery.

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Figure 1: Separable domains of Stn1p interact with Cdc13p and Ten1p.
Figure 2: Stn11–186–Ten1p overproduction makes CDC13 dispensable.
Figure 3: Telomerase requirement for STN1–TEN1-bypass.
Figure 4: A role for DNA replication in alternative capping.

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Acknowledgements

We appreciate helpful input from J. Bachant, A. Grosovsky, D. Lydall, M. Petreaca, R. Wellinger and members of the Nugent laboratory. We thank D. Gottschling, S. Elledge and V. Lundblad for sharing strains and plasmids. We also thank D. Lydall and R. Wellinger for communicating results before publication. This work was funded by a grant from the National Institutes of Health (CIN).

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Author notes

  1. Ruben C. Petreaca and Huan-Chih Chiu: These authors contributed equally to this work.

Authors and Affiliations

  1. Graduate Program in Cell, Molecular and Developmental Biology, 5429 Boyce Hall, University of California Riverside, Riverside, 92521, CA, USA

    Ruben C. Petreaca, Huan-Chih Chiu & Ling Xu

  2. Department of Cell Biology and Neuroscience, 5429 Boyce Hall, University of California Riverside, Riverside, 92521, CA, USA

    Ruben C. Petreaca, Huan-Chih Chiu, Holly A. Eckelhoefer, Charles Chuang, Ling Xu & Constance I. Nugent

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  1. Ruben C. Petreaca
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  6. Constance I. Nugent
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Contributions

R.C.P. made the initial bypass observations and performed the two-hybrid analysis and Southern blots. H.C. analysed ssTG and performed the Pol12-related experiments. R.P., P.C. and H.A.E. contributed experiments for cdc13-1 suppression. H.A.E. and C.C. analysed Rad53 shift and provided technical assistance. C.C. performed the DNA damage sensitivity experiments. L.X. analysed the telomere structure. C.I.N. guided experimental design and helped analyse growth phenotypes and bypass in mutant backgrounds. C.I.N. wrote the paper.

Corresponding author

Correspondence to Constance I. Nugent.

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The authors declare no competing financial interests.

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Supplementary Information

Supplementary Figures S1, S2, S3, S4 and Supplementary Table S1 (PDF 709 kb)

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Petreaca, R., Chiu, HC., Eckelhoefer, H. et al. Chromosome end protection plasticity revealed by Stn1p and Ten1p bypass of Cdc13p. Nat Cell Biol 8, 748–755 (2006). https://doi.org/10.1038/ncb1430

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