Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

PML regulates p53 stability by sequestering Mdm2 to the nucleolus

Nature Cell Biology volume 6, pages 665–672 (2004)Cite this article

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 4–6); 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.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

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.

Similar content being viewed by others

References

  1. Pearson, M. et al. PML regulates p53 acetylation and premature senescence induced by oncogenic Ras. Nature 406, 207–210 (2000).

    Article  CAS  PubMed  Google Scholar 

  2. Guo, A. et al. The function of PML in p53-dependent apoptosis. Nature Cell Biol. 2, 730–736 (2000).

    Article  CAS  PubMed  Google Scholar 

  3. Louria-Hayon, I. et al. PML protects p53 from Mdm2-mediated inhibition and degradation. J. Biol. Chem. 278, 33134–33141 (2003).

    Article  CAS  PubMed  Google Scholar 

  4. Wei, X. et al. Physical and functional interactions between PML and MDM2. J. Biol. Chem. 278, 29288–29297 (2003).

    Article  CAS  PubMed  Google Scholar 

  5. Zhu, H., Wu, L. & Maki, C.G. MDM2 and PML antagonize each other through their direct interaction with p53. J. Biol. Chem. 278, 49286–49292 (2003).

    Article  CAS  PubMed  Google Scholar 

  6. Kurki, S., Latonen, L. & Laiho, M. Cellular stress and DNA damage invoke temporally distinct Mdm2, p53 and PML complexes and damage-specific nuclear relocalization. J. Cell Sci. 116, 3917–3925 (2003).

    Article  CAS  PubMed  Google Scholar 

  7. Oren, M. Regulation of the p53 tumor suppressor protein. J. Biol. Chem. 274, 36031–36034 (1999).

    Article  CAS  PubMed  Google Scholar 

  8. Weber, J.D., Taylor, L.J., Roussel, M.F., Sherr, C.J. & Bar-Sagi, D. Nucleolar Arf sequesters Mdm2 and activates p53. Nature Cell Biol. 1, 20–26 (1999).

    Article  CAS  PubMed  Google Scholar 

  9. Lohrum, M.A., Ludwig, R.L., Kubbutat, M.H., Hanlon, M. & Vousden, K.H. Regulation of HDM2 activity by the ribosomal protein L11. Cancer Cell 3, 577–587 (2003).

    Article  CAS  PubMed  Google Scholar 

  10. Salomoni, P. & Pandolfi, P.P. The role of PML in tumor suppression. Cell 108, 165–170 (2002).

    Article  CAS  PubMed  Google Scholar 

  11. Jensen, K., Shiels, C. & Freemont, P.S. PML protein isoforms and the RBCC/TRIM motif. Oncogene 20, 7223–7233 (2001).

    Article  CAS  PubMed  Google Scholar 

  12. Ferbeyre, G. et al. PML is induced by oncogenic ras and promotes premature senescence. Genes Dev. 14, 2015–2027 (2000).

    CAS  PubMed  PubMed Central  Google Scholar 

  13. Fogal, V. et al. Regulation of p53 activity in nuclear bodies by a specific PML isoform. EMBO J. 19, 6185–6195 (2000).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Rubbi, C.P. & Milner, J. Disruption of the nucleolus mediates stabilization of p53 in response to DNA damage and other stresses. EMBO J. 22, 6068–6077 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Abraham, R.T. Cell-cycle-checkpoint signaling through the ATM and ATR kinases. Genes Dev. 15, 2177–2196 (2001).

    Article  CAS  Google Scholar 

  16. Sarkaria, J.N. et al. Inhibition of ATM and ATR kinase activities by the radiosensitizing agent, caffeine. Cancer Res. 59, 4375–4382 (1999).

    CAS  PubMed  Google Scholar 

  17. Gage, B.M. et al. Spontaneously immortalized cell lines obtained from adult Atm-null mice retain sensitivity to ionizing radiation and exhibit a mutational pattern suggestive of oxidative stress. Oncogene 20, 4291–4297 (2001).

    Article  CAS  PubMed  Google Scholar 

  18. Cliby, W.A. et al. Overexpression of a kinase-inactive ATR protein causes sensitivity to DNA-damaging agents and defects in cell-cycle checkpoints. EMBO J. 17, 159–169 (1998).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Nghiem, P., Park, P.K., Kim, Y., Vaziri, C. & Schreiber, S.L. ATR inhibition selectively sensitizes G1 checkpoint-deficient cells to lethal premature chromatin condensation. Proc. Natl Acad. Sci. USA 98, 9092–9097 (2001).

    Article  CAS  PubMed  Google Scholar 

  20. Bischof O., Kirsh O., Pearson M., Itahana K., Pelicci P.P. & Dejean A. Deconstructing PML-induced premature senescence. EMBO J. 21, 3358–3369 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Stott F.J. et al. The alternative product of the human CDKN2A locus, p14(ARF), participates in a regulatory feedback loop with p53 and MDM2. EMBO J. 17, 5001–5014 (1998).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Zhang, Y. et al. Ribosomal protein L11 negatively regulates oncoprotein MDM2 and mediates a p53-dependent ribosomal-stress checkpoint pathway. Mol. Cell. Biol. 23, 8902–8912 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Carbone, R., Pearson, M., Minucci, S. & Pelicci, P.G. PML NBs associate with the hMre11 complex and p53 at sites of irradiation-induced DNA damage. Oncogene 21, 1633–1640 (2002).

    Article  CAS  PubMed  Google Scholar 

  24. Cliby, W.A., Lewis, K.A., Lilly, K.K. & Kaufmann, S.H. S phase and G2 arrests induced by topoisomerase I poisons are dependent on ATR kinase function. J. Biol. Chem. 277, 1599–1606 (2002).

    Article  CAS  PubMed  Google Scholar 

  25. Yang, S., Kuo, C., Bisi, J.E. & Kim, M.K. PML-dependent apoptosis after DNA damage is regulated by the checkpoint kinase hCds1/Chk2. Nature Cell Biol. 4, 865–870 (2002).

    Article  CAS  PubMed  Google Scholar 

  26. Gostissa, M., Hofmann, T.G., Will, H. & Del Sal, G. Regulation of p53 functions: let's meet at the nuclear bodies. Curr. Opin. Cell Biol. 15, 351–357 (2003).

    Article  CAS  PubMed  Google Scholar 

  27. Bernardi, R. & Pandolfi, P.P. Role of PML and the PML-nuclear body in the control of programmed cell death. Oncogene 22, 9048–9057 (2003).

    Article  CAS  PubMed  Google Scholar 

  28. Andersen, J.S. et al. Directed proteomic analysis of the human nucleolus. Curr. Biol. 12, 1–11 (2002).

    Article  PubMed  Google Scholar 

Download references

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.

Author information

Author notes

  1. Pier Paolo Scaglioni and Stephan Bergmann: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Pathology and Medicine, Cancer Biology and Genetics Program, Sloan-Kettering Institute, Memorial Sloan-Kettering Cancer Center, New York, 10021, NY, USA

    Rosa Bernardi, Pier Paolo Scaglioni, Stephan Bergmann & Pier Paolo Pandolfi

  2. Beatson Institute for Cancer Research, Glasgow, G61 1BD, UK

    Henning F. Horn & Karen H. Vousden

Authors
  1. Rosa Bernardi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Pier Paolo Scaglioni
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Stephan Bergmann
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Henning F. Horn
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Karen H. Vousden
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Pier Paolo Pandolfi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Pier Paolo Pandolfi.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Figures

Figures S1, S2, S3 and S4 (PDF 726 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

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). https://doi.org/10.1038/ncb1147

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ncb1147

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing