Positive feedback between p53 and TRF2 during telomere-damage signalling and cellular senescence

Abstract

The telomere-capping complex shelterin protects functional telomeres and prevents the initiation of unwanted DNA-damage-response pathways. At the end of cellular replicative lifespan, uncapped telomeres lose this protective mechanism and DNA-damage signalling pathways are triggered that activate p53 and thereby induce replicative senescence. Here, we identify a signalling pathway involving p53, Siah1 (a p53-inducible E3 ubiquitin ligase) and TRF2 (telomere repeat binding factor 2; a component of the shelterin complex). Endogenous Siah1 and TRF2 were upregulated and downregulated, respectively, during replicative senescence with activated p53. Experimental manipulation of p53 expression demonstrated that p53 induces Siah1 and represses TRF2 protein levels. The p53-dependent ubiquitylation and proteasomal degradation of TRF2 are attributed to the E3 ligase activity of Siah1. Knockdown of Siah1 stabilized TRF2 and delayed the onset of cellular replicative senescence, suggesting a role for Siah1 and TRF2 in p53-regulated senescence. This study reveals that p53, a downstream effector of telomere-initiated damage signalling, also functions upstream of the shelterin complex.

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Figure 1: Replicative cellular senescence is associated with decreased TRF2 and increased Siah1.
Figure 2: p53 upregulates Siah1 and downregulates TRF2.
Figure 3: Siah1 knockdown stabilizes TRF2.
Figure 4: TRF2 is subject to proteasomal degradation and ubiquitylated in vivo.
Figure 5: Siah1 interacts with, and ubiquitylates, TRF2.
Figure 6: Roles of Siah1 and TRF2 in cellular senescence in vitro and in vivo.

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Acknowledgements

We thank M. Tainsky, B. Vogelstein and T. de Lange for cells and reagents. We also thank K. Kumamoto for carrying out Nutlin-3a treatment, A. Robles for carrying out doxorubicin treatment of lymphoblast cells, M. Yoneda for technical assistance and E. Spillare for continuous support. This research was supported in part by the Intramural Research Program of the NIH, NCI. B.V. was supported by the grants from the Grant Agency of the Czech Republic (number 301/08/1468) and the Internal Grant Agency of Health of Czech Republic (number NS/9812-4). J.C.B. was supported by Breast Cancer Campaign, Cancer-Research UK and the Institut National de la Sante et de la Recherche Medicale. D.L. is a Gibb fellow of Cancer-Research UK.

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K.F., I.H., A.M.M. and L.M.M.J. performed experiments. B.V., J.-C.B. and D.P.L. provided essential reagents and suggestions. K.F., I.H., E.A., L.M.M.J. and C.C.H. coordinated the study and wrote the manuscript. C.C.H. was responsible for the overall project. All authors discussed the results and commented on the manuscript.

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Correspondence to Curtis C. Harris.

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Fujita, K., Horikawa, I., Mondal, A. et al. Positive feedback between p53 and TRF2 during telomere-damage signalling and cellular senescence. Nat Cell Biol 12, 1205–1212 (2010). https://doi.org/10.1038/ncb2123

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