PML regulates p53 stability by sequestering Mdm2 to the nucleolus

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Abstract

The promyelocytic leukaemia (PML) tumour-suppressor protein potentiates p53 function by regulating post-translational modifications, such as CBP-dependent acetylation1,2 and Chk2-dependent phosphorylation, in the PML-Nuclear Body (NB)3. PML was recently shown to interact with the p53 ubiquitin-ligase Mdm2 (refs 46); however, the mechanism by which PML regulates Mdm2 remains unclear. Here, we show that PML enhances p53 stability by sequestering Mdm2 to the nucleolus. We found that after DNA damage, PML and Mdm2 accumulate in the nucleolus in an Arf-independent manner. In addition, we found that the nucleolar localization of PML is dependent on ATR activation and phosphorylation of PML by ATR. Notably, in Pml−/− cells, sequestration of Mdm2 to the nucleolus was impaired, as well as p53 stabilization and the induction of apoptosis. Furthermore, we demonstrate that PML physically associates with the nucleolar protein L11, and that L11 knockdown impairs the ability of PML to localize to nucleoli after DNA damage. These findings demonstrate an unexpected role of PML in the nucleolar network for tumour suppression.

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Figure 1: PML mediates stabilization of p53 in the presence of Mdm2.
Figure 2: PML accumulates in the nucleolus and is phosphorylated by ATR after DNA damage.
Figure 3: Mdm2 and PML colocalize in the nucleolus after DNA damage.
Figure 4: PML interacts directly with Mdm2 and stabilizes p53 through sequestration of Mdm2.
Figure 5: L11 facilitates PML nucleolar localization after DNA damage.

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Acknowledgements

We are indebted to I. Guernah, A. Guo, P. Salomoni, F. Bernassola, and S. Grisendi for suggestions during the course of this work and critical reading of the manuscript. We are grateful to W. Cliby, C. Di Como, W. Gu, A. Levine, S. Lowe, G. Lozano, C. Prives, S. Schreiber, M. Turker, M. van Lohuizen and A. Weismann for reagents and advice. We thank Y. Haupt for useful discussion. R.B. and P.P.S. were supported by T32 training grants from the National Institutes of Health. P.P.S. is also a recipient of an ASCO Young Investigator Award and a CALGB Oncology Fellows Award. This work was supported by the award of a National Institutes of Health grant RO1 CA-71692 to P.P.P.

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Correspondence to Pier Paolo Pandolfi.

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Bernardi, R., Scaglioni, P., Bergmann, S. et al. PML regulates p53 stability by sequestering Mdm2 to the nucleolus. Nat Cell Biol 6, 665–672 (2004) doi:10.1038/ncb1147

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