Mechanical cues control mutant p53 stability through a mevalonate–RhoA axis

Abstract

Tumour-associated p53 missense mutants act as driver oncogenes affecting cancer progression, metastatic potential and drug resistance (gain-of-function)1. Mutant p53 protein stabilization is a prerequisite for gain-of-function manifestation; however, it does not represent an intrinsic property of p53 mutants, but rather requires secondary events2. Moreover, mutant p53 protein levels are often heterogeneous even within the same tumour, raising questions on the mechanisms that control local mutant p53 accumulation in some tumour cells but not in their neighbours2,3. By investigating the cellular pathways that induce protection of mutant p53 from ubiquitin-mediated proteolysis, we found that HDAC6/Hsp90-dependent mutant p53 accumulation is sustained by RhoA geranylgeranylation downstream of the mevalonate pathway, as well as by RhoA- and actin-dependent transduction of mechanical inputs, such as the stiffness of the extracellular environment. Our results provide evidence for an unpredicted layer of mutant p53 regulation that relies on metabolic and mechanical cues.

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Fig. 1: Statins reduce missense mutant p53 protein levels in cancer cells.
Fig. 2: Statins unleash MDM2-mediated degradation of mutant p53 by disrupting its interaction with Hsp90.
Fig. 3: The SREBP–mevalonate pathway controls mutant p53 levels via GGPP.
Fig. 4: RhoA geranylgeranylation controls mutant p53 levels downstream of the mevalonate pathway.
Fig. 5: Mechanical cues control mutant p53 levels and activity via RhoA/actin cytoskeleton.

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Acknowledgements

We thank A. Testa for discussions and proofreading the manuscript. We acknowledge G. Pastore for technical support. We thank S. Giulitti for preparation of hydrogels. We acknowledge support by the Italian Health Ministry (RF-2011-02346976 to G.D.S. and GR-2011-02348707 to D.S.), the Italian University and Research Ministry (PRIN-2015-8KZKE3), the Cariplo Foundation (grant no. 2014-0812) and Beneficentia-Stiftung to G.D.S. This work was supported by grants from the Associazione Italiana per la Ricerca sul Cancro (AIRC) and AIRC Special Program Molecular Clinical Oncology ‘5 per mille’ (grant no. 10016) to G.D.S., S.B., A.R. and S.P., and AIRC IG (grant no. 17659) to G.D.S. This project has received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation programme (grant agreement no. 670126-DENOVOSTEM) and an AIRC PI-Grant and by Epigenetics Flagship project CNR-Miur grants to S.P. M.M. is supported by the FIRB RBAP11Z4Z9 project from the Italian Ministry of Education and the FCT Investigator Programme IF/00694/2013 from the Portuguese Foundation for Science and Technology (FCT), Portugal. R.B. is a fellow of the Fondazione Italiana per la Ricerca sul Cancro (FIRC).

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E.I., G.S., K.L., R.B., A.Z. and L.A. performed the experiments. A.R. performed mouse experiments. M.M. performed the high-content screening. S.B. performed bioinformatic analysis. D.S. and L.U.S. performed AFM experiments. G.S., E.I. and G.D.S. designed experiments. G.S., F.M., S.P. and G.D.S. wrote the manuscript.

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Correspondence to Giannino Del Sal.

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Supplementary Figures 1–6

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Supplementary Table 1

Small molecule screening details

Supplementary Table 2

Small molecule screening results

Supplementary Table 3

List of siRNA sequences used

Supplementary Table 4

List of primers used for qRT-PCR

Supplementary Table 5

Details of the genes composing the stiffness, YAP/TAZ and mutant p53 signatures

Supplementary Table 6

Statistical source data

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Ingallina, E., Sorrentino, G., Bertolio, R. et al. Mechanical cues control mutant p53 stability through a mevalonate–RhoA axis. Nat Cell Biol 20, 28–35 (2018). https://doi.org/10.1038/s41556-017-0009-8

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