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MDMX stability is regulated by p53-induced caspase cleavage in NIH3T3 mouse fibroblasts

Oncogene volume 21, pages 867–877 (2002)Cite this article

Abstract

MDMX is a p53 binding protein, which shares a high degree of homology with MDM2, a negative regulator of the tumor suppressor p53. MDMX has been shown to counteract MDM2-dependent p53 degradation and to stabilize p53 in its inactive form. In this study: we identify two MDMX proteolytic pathways that control its intracellular levels, and show that MDMX post-translational processing may be regulated by p53. Mouse MDMX is cleaved in vitro and in vivo by caspase activity, between aminoacids 358 and 361, producing a p54 minor form. In addition, MDMX is subjected to proteasome-mediated degradation, which concurs to MDMX proteolysis mainly through degradation of p54. A D361A-MDMX mutant, resistant to caspase cleavage, exhibits prolonged intracellular lifetime in comparison to wild-type protein, indicating that caspase cleavage affects stability of MDMX protein probably by modulating its further degradation. Overexpression of exogenous p53 increases the intracellular levels of p54 product. Similarly, activation of endogenous p53 by adriamycin enhances MDMX cleavage and produces a marked decrease of its intracellular levels, while not affecting the D361A-MDMX mutant. In addition, the D361A-MDMX mutant lacks the ability to inhibit p53 transactivation in respect to wild-type MDMX, suggesting that MDMX caspase cleavage play an important functional role. In conclusion, our results demonstrate that, in analogy to MDM2, MDMX may be subjected to proteolytic modifications that regulate its intracellular levels. Moreover, decrease of MDMX protein levels following p53 activation suggests a p53-dependent regulatory feedback of MDMX function.

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Acknowledgements

We thank M Oren for the gift of MDM2 plasmid and AJ Levine for anti-MDM2 antibodies. We are grateful to Silvia Soddu (Regina Elena Cancer Institute, Rome, Italy) for helpful discussion. This work was supported by research grants from Ministero dell'Università e della Ricerca Scientifica e Tecnologica (MURST), Ministero della Sanità, Progetto Italia-USA and Associazione Italiana per la Ricerca sul Cancro (AIRC).

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Authors and Affiliations

  1. Molecular Oncogenesis Laboratory, Regina Elena Cancer Institute, Via delle Messi D'Oro 156, 00158 Rome, Italy

    Francesca Gentiletti, Francesca Mancini, Marco D'Angelo, Ada Sacchi, Alfredo Pontecorvi & Fabiola Moretti

  2. Medical Pathology Institute, Catholic University, Largo Vito 1, 00100 Rome, Italy

    Alfredo Pontecorvi

  3. Department of Molecular and Cell Biology, Leiden University Medical Center, PO Box 9503, 2300 RA Leiden, The Netherlands

    Aart G Jochemsen

  4. Neurobiology and Molecular Medicine Institute, National Council of Research, Viale Kant 15, 00156 Rome, Italy

    Fabiola Moretti

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  1. Francesca Gentiletti
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Correspondence to Fabiola Moretti.

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Gentiletti, F., Mancini, F., D'Angelo, M. et al. MDMX stability is regulated by p53-induced caspase cleavage in NIH3T3 mouse fibroblasts. Oncogene 21, 867–877 (2002). https://doi.org/10.1038/sj.onc.1205137

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