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Hsp70 molecular chaperones are required to support p53 tumor suppressor activity under stress conditions

Oncogene volume 28, pages 4284–4294 (2009)Cite this article

Abstract

p53 as an unstable protein in vitro likely requires stabilizing factors to act as a tumor suppressor in vivo. Here, we show that in human cells transfected with wild-type (WT) p53, Hsp90 and Hsp70 molecular chaperones maintain the p53 native conformation under heat-shock conditions (42 °C) as well as assist p53 refolding at 37 °C, during the recovery from heat shock. We also show that the interaction of WT p53 with WAF1 promoter in cells is sensitive to Hsp70 and Hsp90 inhibition already at 37 °C and further decreased on heat shock. The influence of chaperones on p53 binding to the WAF1 promoter sequence has been confirmed in vitro, using highly purified proteins. Hsp90 stabilizes the binding of p53 to the promoter sequence at 37 °C, whereas under heat-shock conditions the requirement for the Hsp70-Hsp40 system and its cooperation with Hsp90 increases. Hop co-chaperone additionally stimulates these reactions. Interestingly, the combined Hsp90 and Hsp70-Hsp40 allow for a limited in vitro restoration of the DNA-binding activity by the p53 oncogenic variant R249S and affect its conformation in cells. Our results indicate for the first time that, especially under stress conditions, not only Hsp90 but also Hsp70 is required for the chaperoning of WT and R249S p53.

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Acknowledgements

We thank Dr Marcin Klejman for sharing purified Hsp90β proteins; Professor Peter Csermely for the plasmid encoding human Hsp90α; Professor Ted Hupp for plasmid for p53 overexpression; Professor Richard Morimoto for plasmids encoding human Hsp70, Hdj1 and Hop; and Professor Kazutoyo Terada for construct for the Hdj2 overexpression. Dawid Walerych is the recipient of a scholarship from the Postgraduate School of Molecular Medicine affiliated with the Medical University of Warsaw. This work was supported by a grant from the Polish Ministry of Science and Higher Education.

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Author notes

  1. D Walerych and M B Olszewski: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Molecular Biology, International Institute of Molecular and Cell Biology in Warsaw, Warsaw, Poland

    D Walerych, M B Olszewski, M Gutkowska, A Helwak, M Zylicz & A Zylicz

  2. The Nencki Institute of Experimental Biology PAS, Warsaw, Poland

    M Gutkowska

  3. The Institute of Biochemistry and Biophysics PAS, Warsaw, Poland

    A Helwak

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  1. D Walerych
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Correspondence to M Zylicz or A Zylicz.

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

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Walerych, D., Olszewski, M., Gutkowska, M. et al. Hsp70 molecular chaperones are required to support p53 tumor suppressor activity under stress conditions. Oncogene 28, 4284–4294 (2009). https://doi.org/10.1038/onc.2009.281

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