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Transcription-independent ARF regulation in oncogenic stress-mediated p53 responses

Nature volume 464, pages 624–627 (2010)Cite this article

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Abstract

The tumour suppressor ARF is specifically required for p53 activation under oncogenic stress1,2,3,4,5,6. Recent studies showed that p53 activation mediated by ARF, but not that induced by DNA damage, acts as a major protection against tumorigenesis in vivo under certain biological settings7,8, suggesting that the ARF–p53 axis has more fundamental functions in tumour suppression than originally thought. Because ARF is a very stable protein in most human cell lines, it has been widely assumed that ARF induction is mediated mainly at the transcriptional level and that activation of the ARF–p53 pathway by oncogenes is a much slower and largely irreversible process by comparison with p53 activation after DNA damage. Here we report that ARF is very unstable in normal human cells but that its degradation is inhibited in cancerous cells. Through biochemical purification, we identified a specific ubiquitin ligase for ARF and named it ULF. ULF interacts with ARF both in vitro and in vivo and promotes the lysine-independent ubiquitylation and degradation of ARF. ULF knockdown stabilizes ARF in normal human cells, triggering ARF-dependent, p53-mediated growth arrest. Moreover, nucleophosmin (NPM) and c-Myc, both of which are commonly overexpressed in cancer cells, are capable of abrogating ULF-mediated ARF ubiquitylation through distinct mechanisms, and thereby promote ARF stabilization in cancer cells. These findings reveal the dynamic feature of the ARF–p53 pathway and suggest that transcription-independent mechanisms are critically involved in ARF regulation during responses to oncogenic stress.

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Figure 1: ULF is identified as a major factor for short half-lives of ARF in normal human fibroblast cells.
Figure 2: ULF interacts with ARF.
Figure 3: ULF-mediated effect on ARF ubiquitylation and degradation is modulated by NPM.
Figure 4: c-Myc overexpression blocks the interaction between ULF and ARF, which leads to c-Myc-mediated, transcription-independent ARF induction.

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Data deposits

The full-length ULF sequence is deposited in GenBank under accession number EU489742.

References

  1. Lowe, S. W. & Sherr, C. J. Tumor suppression by Ink4a-Arf: progress and puzzles. Curr. Opin. Genet. Dev. 13, 77–83 (2003)

    Article  CAS  PubMed  Google Scholar 

  2. Gil, J. & Peters, G. Regulation of the INK4b-ARF-INK4a tumour suppressor locus: all for one or one for all. Nature Rev. Mol. Cell Biol. 7, 667–677 (2006)

    Article  CAS  Google Scholar 

  3. Matheu, A., Maraver, A. & Serrano, M. The Arf/p53 pathway in cancer and aging. Cancer Res. 68, 6031–6034 (2008)

    Article  CAS  PubMed  Google Scholar 

  4. Brooks, C. L. & Gu, W. p53 ubiquitination: Mdm2 and beyond. Mol. Cell 21, 307–315 (2006)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Sherr, C. J. Divorcing ARF and p53: an unsettled case. Nature Rev. Cancer 6, 663–673 (2006)

    Article  CAS  Google Scholar 

  6. Kruse, J.-P. & Gu, W. Modes of p53 regulation. Cell 137, 609–622 (2009)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Christophorou, M. A., Ringshausen, I., Finch, A. J., Swigart, L. B. & Evan, G. I. The pathological response to DNA damage does not contribute to p53-mediated tumour suppression. Nature 443, 214–217 (2006)

    Article  ADS  CAS  PubMed  Google Scholar 

  8. Martins, C. P., Brown-Swigart, L. & Evan, G. I. Modeling the therapeutic efficacy of p53 restoration in tumors. Cell 127, 1323–1334 (2006)

    Article  CAS  PubMed  Google Scholar 

  9. Kuo, M. L., den Besten, W., Bertwistle, D., Roussel, M. F. & Sherr, C. J. N-terminal polyubiquitination and degradation of the Arf tumor suppressor. Genes Dev. 18, 1862–1874 (2004)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Itahana, K. et al. Tumor suppressor ARF degrades B23, a nucleolar protein involved in ribosome biogenesis and cell proliferation. Mol. Cell 12, 1151–1164 (2003)

    Article  CAS  PubMed  Google Scholar 

  11. Korgaonkar, C. et al. Nucleophosmin (B23) targets ARF to nucleoli and inhibits its function. Mol. Cell. Biol. 25, 1258–1271 (2005)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Brady, S. N., Yu, Y., Maggi, L. B. & Weber, J. D. ARF impedes NPM/B23 shuttling in an Mdm2-sensitive tumor suppressor pathway. Mol. Cell. Biol. 24, 9327–9338 (2004)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Bertwistle, D., Sugimoto, M. & Sherr, C. J. Physical and functional interactions of the Arf tumor suppressor protein with nucleophosmin/B23. Mol. Cell. Biol. 24, 985–996 (2004)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Chen, D. L. et al. ARF-BP1/mule is a critical mediator of the ARF tumor suppressor. Cell 121, 1071–1083 (2005)

    Article  CAS  PubMed  Google Scholar 

  15. Moulin, S., Llanos, S., Kim, S. H. & Peters, G. Binding to nucleophosmin determines the localization of human and chicken ARF but not its impact on p53. Oncogene 27, 2382–2389 (2008)

    Article  CAS  PubMed  Google Scholar 

  16. Colombo, E. et al. Nucleophosmin is required for DNA integrity and p19Arf protein stability. Mol. Cell. Biol. 25, 8874–8886 (2005)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Grisendi, S. et al. Role of nucleophosmin in embryonic development and tumorigenesis. Nature 437, 147–153 (2005)

    Article  ADS  CAS  PubMed  Google Scholar 

  18. Lee, J. W., Choi, H. S., Gyuris, J., Brent, R. & Moore, D. D. Two classes of proteins dependent on either the presence or absence of thyroid hormone for interaction with the thyroid hormone receptor. Mol. Endocrinol. 9, 243–254 (1995)

    CAS  PubMed  Google Scholar 

  19. den Besten, W., Kuo, M. L., Williams, R. T. & Sherr, C. J. Myeloid leukemia-associated nucleophosmin mutants perturb p53-dependent and independent activities of the Arf tumor suppressor protein. Cell Cycle 4, 1593–1598 (2005)

    Article  CAS  PubMed  Google Scholar 

  20. Colombo, E. et al. Delocalization and destabilization of the Arf tumor suppressor by the leukemia-associated NPM mutant. Cancer Res. 66, 3044–3050 (2006)

    Article  CAS  PubMed  Google Scholar 

  21. Cheng, K. et al. The leukemia-associated cytoplasmic nucleophosmin mutant is an oncogene with paradoxical functions: Arf inactivation and induction of cellular senescence. Oncogene 26, 7391–7400 (2007)

    Article  CAS  PubMed  Google Scholar 

  22. Feuerstein, N. & Mond, J. J. ‘Numatrin,’ a nuclear matrix protein associated with induction of proliferation in B lymphocytes. J. Biol. Chem. 262, 11389–11397 (1987)

    CAS  PubMed  Google Scholar 

  23. Nozawa, Y., vanBelzen, N., vanderMade, A. C. J., Dinjens, W. N. M. & Bosman, F. T. Expression of nucleophosmin/B23 in normal and neoplastic colorectal mucosa. J. Pathol. 178, 48–52 (1996)

    Article  CAS  PubMed  Google Scholar 

  24. Falini, B. et al. Cytoplasmic nucleophosmin in acute myelogenous leukemia with a normal karyotype. N. Engl. J. Med. 352, 254–266 (2005)

    Article  CAS  PubMed  Google Scholar 

  25. Quentmeier, H. et al. Cell line OCI/AML3 bears exon-12 NPM gene mutation-A and cytoplasmic expression of nucleophosmin. Leukemia 19, 1760–1767 (2005)

    Article  CAS  PubMed  Google Scholar 

  26. Grisendi, S., Mecucci, C., Falini, B. & Pandolfi, P. P. Nucleophosmin and cancer. Nature Rev. Cancer 6, 493–505 (2006)

    Article  CAS  Google Scholar 

  27. Saporita, A. J., Maggi, L. B., Apicelli, A. J. & Weber, J. D. Therapeutic targets in the ARF tumor suppressor pathway. Curr. Med. Chem. 14, 1815–1827 (2007)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Finak, G. et al. Stromal gene expression predicts clinical outcome in breast cancer. Nature Med. 14, 518–527 (2008)

    Article  CAS  PubMed  Google Scholar 

  29. Buchholz, M. et al. Transcriptome analysis of microdissected pancreatic intraepithelial neoplastic lesions. Oncogene 44, 6626–6636 (2005)

    Article  Google Scholar 

  30. Zindy, F. et al. Myc signaling via the ARF tumor suppressor regulates p53-dependent apoptosis and immortalization. Genes Dev. 12, 2424–2433 (1998)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Abide, W. M. & Gu, W. p53-Dependent and p53-independent activation of autophagy by ARF. Cancer Res. 68, 352–357 (2006)

    Article  Google Scholar 

  32. Li, M. et al. Mono- versus polyubiquitination: differential control of p53 fate by Mdm2. Science 302, 1972–1975 (2003)

    Article  ADS  CAS  PubMed  Google Scholar 

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Acknowledgements

We thank R. Baer and R. Dalla-Favera for critical comments on this study; B. Falini and J. B. Yoon for providing reagents; W. Z. Zhang for helping in mass spectrometry analysis; E. McIntush for developing the anti-ULF antibody. This study was supported by grants from National Institutes of Health/National Cancer Institute, the Leukemia and Lymphoma Society and NSFC-30628028. W.G. is an Ellison Medical Foundation Senior Scholar in aging.

Author Contributions The experiments were conceived and designed by D.C. and W.G. Experiments were performed mainly by D.C. Protein identification, mass spectrometric analysis and cloning were performed by D.C., J.S., W.Z. and J.Q. The paper was written by D.C. and W.G.

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Authors and Affiliations

  1. and Department of Pathology and Cell Biology College of Physicians & Surgeons, Institute for Cancer Genetics, Columbia University, 1130 St Nicholas Avenue, New York, New York 10032, USA,

    Delin Chen, Jing Shan & Wei Gu

  2. Department of Biochemistry and Molecular Biology, Peking University Health Science Center, Beijing 100191, China

    Wei-Guo Zhu

  3. Departments of Biochemistry and Cell Biology, Baylor College of Medicine, One Baylor Plaza, Houston, Texas 77030, USA,

    Jun Qin

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Correspondence to Wei Gu.

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Chen, D., Shan, J., Zhu, WG. et al. Transcription-independent ARF regulation in oncogenic stress-mediated p53 responses. Nature 464, 624–627 (2010). https://doi.org/10.1038/nature08820

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