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A single-codon mutation converts HPV16 E6 oncoprotein into a potential tumor suppressor, which induces p53-dependent senescence of HPV-positive HeLa cervical cancer cells

Oncogene volume 28, pages 762–772 (2009)Cite this article

Abstract

High-risk mucosal human papillomaviruses (HPV), mainly HPV16 and HPV18, are implicated in cervical carcinogenesis. HPV16 E6 oncoprotein binds and often targets for degradation numerous cell proteins, including the tumor suppressor p53 and several PDZ domain proteins. Here, we show that a single-point mutation, F47R, is sufficient to convert the HPV16 E6 oncoprotein into a suppressor of HPV-positive HeLa cervical cancer cells proliferation. The E6 F47R mutant is defective for polyubiquitination and subsequent degradation of p53. When expressed in HPV-positive cervical cancer cells, E6 F47R acts as a dominant negative mutant by counteracting the p53 degradation activity of endogenous E6 and restoring high p53 protein levels. Moreover, the prolonged expression of E6 F47R leads to suppression of HeLa cells proliferation through the induction of premature senescence. This phenotype is independent on the PDZ-binding activity of E6. F47R-senescent HeLa cells exhibit a sustained expression of p53, hMDM2 and p21CIP proteins and a reduced expression of endogenous HPV18 E6 protein. Finally, small interfering RNAs directed against p53 counteract the effect of E6 F47R expression, indicating that E6 F47R-induced cellular senescence is strongly dependent on p53 signaling pathway.

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Acknowledgements

We thank F Hoppe-Seyler for providing HeLa cells, H Zentgraf for the antibody 18E7C, G de Murcia for the PARP antibody and Arbor Vita Corp. (USA) for the HPV18 E6 antibody. We thank E Weiss, A-P Sibler, A-S Rinaldi and B Chatton for helpful discussions. This study was supported by the University of Strasbourg, CNRS, Association de Recherche contre le Cancer (ARC), Ligue Nationale Contre le Cancer. TR and SF were supported by the Canceropôle Grand-EST (INCA).

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  1. Equipe Oncoprotéines, CNRS-UMR 7175, ESBS, Boulevard Sébastien Brant, Illkirch, Bas-Rhin, France

    T Ristriani, S Fournane, G Orfanoudakis, G Travé & M Masson

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  1. T Ristriani
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  2. S Fournane
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  3. G Orfanoudakis
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  4. G Travé
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  5. M Masson
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Correspondence to G Travé or M Masson.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc)

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Ristriani, T., Fournane, S., Orfanoudakis, G. et al. A single-codon mutation converts HPV16 E6 oncoprotein into a potential tumor suppressor, which induces p53-dependent senescence of HPV-positive HeLa cervical cancer cells. Oncogene 28, 762–772 (2009). https://doi.org/10.1038/onc.2008.422

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