Amplification of PPM1D in human tumors abrogates p53 tumor-suppressor activity

Abstract

Expression of oncogenic Ras in primary human cells activates p53, thereby protecting cells from transformation. We show that in Ras-expressing IMR-90 cells, p53 is phosphorylated at Ser33 and Ser46 by the p38 mitogen-activated protein kinase (MAPK). Activity of p38 MAPK is regulated by the p53-inducible phosphatase PPM1D, creating a potential feedback loop. Expression of oncogenic Ras suppresses PPM1D mRNA induction, leaving p53 phosphorylated at Ser33 and Ser46 and in an active state. Retrovirus-mediated overexpression of PPM1D reduced p53 phosphorylation at these sites, abrogated Ras-induced apoptosis and partially rescued cells from cell-cycle arrest. Inactivation of p38 MAPK (the product of Mapk14) in vivo by gene targeting or by PPM1D overexpression expedited tumor formation after injection of mouse embryo fibroblasts (MEFs) expressing E1A+Ras into nude mice. The gene encoding PPM1D (PPM1D, at 17q22/q23) is amplified in human breast-tumor cell lines and in approximately 11% of primary breast tumors, most of which harbor wildtype p53. These findings suggest that inactivation of the p38 MAPK through PPM1D overexpression resulting from PPM1D amplification contributes to the development of human cancers by suppressing p53 activation.

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Figure 1: Human p53 is phosphorylated at Ser33 and Ser46 after H-rasV12 infection.
Figure 2: Overexpression of PPM1D abrogates apoptosis and partially reverses H-rasV12–induced cell-cycle arrest in IMR-90 cells.
Figure 3: PPM1D phosphatase functions in the p38 MAPK/p53 pathway to regulate transformation in vivo.
Figure 4: PPM1D amplification, messenger RNA accumulation and p38 MAPK activation.

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Acknowledgements

We are grateful to J. Hildesheim for help with real-time PCR, J. Clark for help with DNA sequencing and T. Dennis for help with FISH analysis. This study was supported in part by a US Army Breast Cancer Idea Award (to C.W.A.) at the Brookhaven National Laboratory under contract with the US Department of Energy.

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Correspondence to Ettore Appella.

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The authors declare no competing financial interests.

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Bulavin, D., Demidov, O., Saito, S. et al. Amplification of PPM1D in human tumors abrogates p53 tumor-suppressor activity. Nat Genet 31, 210–215 (2002). https://doi.org/10.1038/ng894

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