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Erosion of the telomeric single-strand overhang at replicative senescence

Nature Genetics volume 33, pages 492–496 (2003)Cite this article

Abstract

Cultured primary human cells inevitably enter a state of replicative senescence for which the specific molecular trigger is unknown. We show that the single-strand telomeric overhang, a key component of telomere structure, is eroded at senescence. Expression of telomerase prevents overhang loss, suggesting that this enzyme prevents senescence by maintaining proper telomere structure. In contrast, progressive overhang loss occurs in cells that avoid senescence through the inactivation of p53 and Rb, indicating that overhang erosion is the result of continuous cell division and not a consequence of senescence. We thus provide evidence for a specific molecular alteration in telomere structure at senescence and suggest that this change, rather than overall telomere length, serves to trigger this state.

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Figure 1: Measurement of the telomeric overhang using T-OLA.
Figure 2: Loss of the telomeric overhang at replicative senescence.
Figure 3: Loss of the telomeric overhang in cells expressing large T-antigen.
Figure 4: Effect of growth arrest and physiologic stress on telomeric overhang length.

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Acknowledgements

We would like to thank E. D'Ambrosio for helpful advice regarding T-OLA and for sharing unpublished results, F. Barany for repeated helpful advice, T. de Lange and J. Griffith for plasmids and K. Yates, Y. Dor, J. Yang, R. Watnick, P. Gupta, H. Vaziri, S. Dessain and other members of R.A.W.'s laboratory for helpful comments, discussion and critical review of the manuscript. This work was supported by Merck/MIT (R.A.W.), the US National Cancer Institute (R.A.W.), an American Association for Cancer Research Postdoctoral Fellowship (S.A.S.), an EMBO Long-Term Fellowship (I.B.), the Dana Farber Cancer Institute (V.J.C.), the US National Cancer Institute Howard Temin award (W.C.H.), a Doris Duke Charitable Foundation Clinical Scientist Development Award (W.C.H.) and a Kimmel Scholar Award (W.C.H.). R.A.W. is an American Cancer Society Research Professor and a Daniel K. Ludwig Cancer Research Professor.

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Author notes

  1. Sheila A. Stewart and Ittai Ben-Porath: These authors contributed equally to this work.

Authors and Affiliations

  1. Whitehead Institute for Biomedical Research, Cambridge, 02142, Massachusetts, USA

    Sheila A. Stewart, Ittai Ben-Porath, Benjamin F. O'Connor & Robert A. Weinberg

  2. Channing Laboratory, Dana-Farber Cancer Institute, Brigham and Women's Hospital, Harvard Medical School, Boston, 02115, Massachusetts, USA

    Vincent J. Carey

  3. Departments of Medical Oncology and Medicine, Dana-Farber Cancer Institute, Brigham and Women's Hospital, Harvard Medical School, Boston, 02115, Massachusetts, USA

    William C. Hahn

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  1. Sheila A. Stewart
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Correspondence to Robert A. Weinberg.

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Stewart, S., Ben-Porath, I., Carey, V. et al. Erosion of the telomeric single-strand overhang at replicative senescence. Nat Genet 33, 492–496 (2003). https://doi.org/10.1038/ng1127

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