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Regulation of autophagy by cytoplasmic p53

Abstract

Multiple cellular stressors, including activation of the tumour suppressor p53, can stimulate autophagy. Here we show that deletion, depletion or inhibition of p53 can induce autophagy in human, mouse and nematode cells subjected to knockout, knockdown or pharmacological inhibition of p53. Enhanced autophagy improved the survival of p53-deficient cancer cells under conditions of hypoxia and nutrient depletion, allowing them to maintain high ATP levels. Inhibition of p53 led to autophagy in enucleated cells, and cytoplasmic, not nuclear, p53 was able to repress the enhanced autophagy of p53−/− cells. Many different inducers of autophagy (for example, starvation, rapamycin and toxins affecting the endoplasmic reticulum) stimulated proteasome-mediated degradation of p53 through a pathway relying on the E3 ubiquitin ligase HDM2. Inhibition of p53 degradation prevented the activation of autophagy in several cell lines, in response to several distinct stimuli. These results provide evidence of a key signalling pathway that links autophagy to the cancer-associated dysregulation of p53.

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Figure 1: Induction of autophagic vacuolization by deletion, depletion or inhibition of p53.
Figure 2: PFT-α triggers autophagic vacuolization and has no effect on vacuole-lysosome fusion.
Figure 3: Role of AMPK, p70S6K and mTOR in the p53-mediated modulation of autophagy.
Figure 4: p53 deletion attenuates ATP depletion during glucose deprivation and favours survival under metabolic stress.
Figure 5: Inhibition of autophagy by cytoplasmic p53.
Figure 6: p53 inhibition induces autophagy by ER stress (ac) WT HCT116 cells were transiently transfected with GFP–LC3, treated with PFT-α for 2, 4 or 6 h and subjected to immunofluorescence staining to observe the colocalization of GFP–LC3+puncta with the ER marker ERp57 (a) or a mitochondrial marker (b).
Figure 7: Induction of autophagy requires p53 degradation mediated by HDM2 and the proteasome.

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Acknowledgements

We thank N. Mizushima for GFP–LC3-transgenic mice, Levine and B. Vogelstein for cell lines, J. Chipuk, C. G. Maki, M. Oren and K. Vousden for mutant p53 plasmids, E. Zaharioudaki, B. Gardie-Capdeville, C. Ladrou, M.R. Duchen, A. Jalil, F. Fanelli and A. Petrini for expert assistance. The cep-1(gk138) mutant strain (TJ1) was provided by the Caenorhabditis Genetics Center, which is funded by the NIH National Center for Research Resources (NCRR). N.T. is funded by EMBO and the EU Sixth Framework Programme. F.C. is funded by Fondazione Telethon, AIRC, Compagnia di San Paolo and the Italian Ministry of Health. E.T. is a recipient of a PhD fellowship from Institut National contre le Cancer (INCa). G.K. is supported by Ligue Nationale contre le Cancer, Agence Nationale pour la Recherche, Cancéropôle Ile-de-France, INCa, Fondation pour la Recherche Médicale, and European Union (Active p53, Apo-Sys, ApopTrain, ChemoRes, TransDeath, RIGHT).

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Authors

Contributions

E.T., M.C.M., L.G. and I.V. conducted experiments, prepared figures and analysed data; M.D.-M., M.D.'A., A.C., E.M., C.Z., F.H., U.N., C.S., P.P., J.M.V, R.C., F.M., P.P.B, G.S., G.P., K.B., N.T., P.C. and F.C. performed experiments; E.T. and G.K. planned the project; G.K. supervised the project and wrote the manuscript.

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Correspondence to Guido Kroemer.

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Tasdemir, E., Maiuri, M., Galluzzi, L. et al. Regulation of autophagy by cytoplasmic p53. Nat Cell Biol 10, 676–687 (2008). https://doi.org/10.1038/ncb1730

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