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HDM2 antagonist Nutlin-3 disrupts p73-HDM2 binding and enhances p73 function

Oncogene volume 27, pages 997–1003 (2008)Cite this article

Abstract

Nutlin-3, a small molecule inhibitor, activates p53 by disrupting p53-HDM2 association. In this study, we found that Nutlin-3 suppressed cell growth and induced apoptosis in the absence of wild-type p53, suggesting a p53-independent mechanism for Nutlin-3-induced cell death. Like p53, its homolog p73 transactivates proapoptotic genes and induces cell death. Since HDM2, a key negative regulator of p53, also binds to and inhibits p73, we asked whether p73 could mediate Nutlin-3-induced apoptosis. We demonstrate that Nutlin-3 inhibits endogenous binding between the proapoptotic p73 isoform TAp73α and HDM2 in p53-null cells. Dissociation of p73 and HDM2 leads to increased p73 transcriptional activity with upregulation of p73 target genes noxa, puma and p21, as well as enhanced apoptosis. p73 knockdown by siRNA results in rescue of Nutlin-3-treated cells, indicating that Nutlin-3-induced apoptosis is, at least in part, p73 dependent. In addition, Nutlin-3 treatment increases TAp73α protein levels with prolongation of p73 half-life. These results provide the first evidence that Nutlin-3 disrupts endogenous p73–HDM2 interaction and enhances the stability and proapoptotic activities of p73 and thus, provides a rationale for the use of Nutlin-3 in the large number of human tumors in which p53 is inactivated.

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Acknowledgements

We thank Lynn Cheng for technical assistance and Ian Watson and members of the Irwin Lab. This project is supported by The James Fund for Neuroblastoma Research at SickKids, the Terry Fox Foundation and the Canadian Cancer Society through the National Cancer Institute of Canada (NCIC). LL is a research fellow of the Terry Fox Foundation through the NCIC. MSI is Canada Research chair in Cancer Biology.

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Authors and Affiliations

  1. Division of Hematology/Oncology, Department of Paediatrics, Hospital for Sick Children, University of Toronto, Ontario, Canada

    L M S Lau, X Zhao & M S Irwin

  2. Cell Biology Program, Research Institute, Hospital for Sick Children, University of Toronto, Ontario, Canada

    L M S Lau, J K Nugent & M S Irwin

  3. Institute of Medical Sciences and Department of Laboratory Medicine and Pathobiology, University of Toronto, Ontario, Canada

    M S Irwin

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  1. L M S Lau
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  2. J K Nugent
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  4. M S Irwin
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Correspondence to M S Irwin.

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

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Lau, L., Nugent, J., Zhao, X. et al. HDM2 antagonist Nutlin-3 disrupts p73-HDM2 binding and enhances p73 function. Oncogene 27, 997–1003 (2008). https://doi.org/10.1038/sj.onc.1210707

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