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The ubiquitin ligase COP1 is a critical negative regulator of p53

Nature volume 429, pages 86–92 (2004)Cite this article

Abstract

COP1 (constitutively photomorphogenic 1) is a RING-finger-containing protein that functions to repress plant photomorphogenesis, the light-mediated programme of plant development. Mutants of COP1 are constitutively photomorphogenic, and this has been attributed to their inability to negatively regulate the proteins LAF1 (ref. 1) and HY5 (ref. 2). The role of COP1 in mammalian cells is less well characterized3. Here we identify the tumour-suppressor protein p53 as a COP1-interacting protein. COP1 increases p53 turnover by targeting it for degradation by the proteasome in a ubiquitin-dependent fashion, independently of MDM2 or Pirh2, which are known to interact with and negatively regulate p53. Moreover, COP1 serves as an E3 ubiquitin ligase for p53 in vitro and in vivo, and inhibits p53-dependent transcription and apoptosis. Depletion of COP1 by short interfering RNA (siRNA) stabilizes p53 and arrests cells in the G1 phase of the cell cycle. Furthermore, we identify COP1 as a p53-inducible gene, and show that the depletion of COP1 and MDM2 by siRNA cooperatively sensitizes U2-OS cells to ionizing-radiation-induced cell death. Overall, these results indicate that COP1 is a critical negative regulator of p53 and represents a new pathway for maintaining p53 at low levels in unstressed cells.

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Figure 1: COP1 and its interaction with p53.
Figure 2: COP1 negative regulation of p53.
Figure 3: siRNA ablation of COP1 causes an accumulation of p53 protein and induces a G1 arrest.
Figure 4: COP1, Pirh2 and MDM2 ablation by siRNA stabilizes and activates p53.
Figure 5: COP1 is a p53-inducible gene.

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References

  1. Seo, H. S. et al. LAF1 ubiquitination by COP1 controls photomorphogenesis and is stimulated by SPA1. Nature 424, 995–999 (2003)

    Article  ADS  Google Scholar 

  2. Ang, L. H. et al. Molecular interaction between COP1 and HY5 defines a regulatory switch for light control of Arabidopsis development. Mol. Cell 1, 213–222 (1998)

    Article  CAS  Google Scholar 

  3. Yi, C., Wang, H., Wei, N. & Deng, X. W. An initial biochemical and cell biological characterization of the mammalian homologue of a central plant developmental switch, COP1. BMC Cell Biol. 3, 30 (2002)

    Article  Google Scholar 

  4. Neff, M. M., Fankhauser, C. & Chory, J. Light: an indicator of time and place. Genes Dev. 14, 257–271 (2000)

    CAS  Google Scholar 

  5. Ma, L. et al. Light control of Arabidopsis development entails coordinated regulation of genome expression and cellular pathways. Plant Cell 13, 2589–2607 (2001)

    Article  CAS  Google Scholar 

  6. Ma, L. et al. Genomic evidence for COP1 as a repressor of light-regulated gene expression and development in Arabidopsis. Plant Cell 14, 2383–2398 (2002)

    Article  CAS  Google Scholar 

  7. Hardtke, C. S. & Deng, X. W. The cell biology of the COP/DET/FUS proteins. Regulating proteolysis in photomorphogenesis and beyond? Plant Physiol. 124, 1548–1557 (2000)

    Article  CAS  Google Scholar 

  8. Bianchi, E. et al. Characterization of human constitutive photomorphogenesis protein 1, a RING finger ubiquitin ligase that interacts with Jun transcription factors and modulates their transcriptional activity. J. Biol. Chem. 278, 19682–19690 (2003)

    Article  CAS  Google Scholar 

  9. Oren, M. Decision making by p53: life, death and cancer. Cell Death Differ. 10, 431–442 (2003)

    Article  CAS  Google Scholar 

  10. Jin, S. & Levine, A. J. The p53 functional circuit. J. Cell Sci. 114, 4139–4140 (2001)

    CAS  Google Scholar 

  11. Shimizu, H. & Hupp, T. R. Intrasteric regulation of MDM2. Trends Biochem. Sci. 28, 346–349 (2003)

    Article  CAS  Google Scholar 

  12. Honda, R., Tanaka, H. & Yasuda, H. Oncoprotein MDM2 is a ubiquitin ligase E3 for tumor suppressor p53. FEBS Lett. 420, 25–27 (1997)

    Article  CAS  Google Scholar 

  13. Kubbutat, M. H., Jones, S. N. & Vousden, K. H. Regulation of p53 stability by Mdm2. Nature 387, 299–303 (1997)

    Article  ADS  CAS  Google Scholar 

  14. Haupt, Y., Maya, R., Kazaz, A. & Oren, M. Mdm2 promotes the rapid degradation of p53. Nature 387, 296–299 (1997)

    Article  ADS  CAS  Google Scholar 

  15. Leng, R. P. et al. Pirh2, a p53-induced ubiquitin-protein ligase, promotes p53 degradation. Cell 112, 779–791 (2003)

    Article  CAS  Google Scholar 

  16. Thut, C. J., Goodrich, J. A. & Tjian, R. Repression of p53-mediated transcription by MDM2: a dual mechanism. Genes Dev. 11, 1974–1986 (1997)

    Article  CAS  Google Scholar 

  17. Mirnezami, A. H. et al. Hdm2 recruits a hypoxia-sensitive corepressor to negatively regulate p53-dependent transcription. Curr. Biol. 13, 1234–1239 (2003)

    Article  CAS  Google Scholar 

  18. Allan, L. A. & Fried, M. p53-dependent apoptosis or growth arrest induced by different forms of radiation in U2OS cells: p21WAF1/CIP1 repression in UV induced apoptosis. Oncogene 18, 5403–5412 (1999)

    Article  CAS  Google Scholar 

  19. Qian, H., Wang, T., Naumovski, L., Lopez, C. D. & Brachmann, R. K. Groups of p53 target genes involved in specific p53 downstream effects cluster into different classes of DNA binding sites. Oncogene 21, 7901–7911 (2002)

    Article  CAS  Google Scholar 

  20. Doonan, J. & Hunt, T. Cell cycle Why don't plants get cancer? Nature 380, 481–482 (1996)

    Article  ADS  CAS  Google Scholar 

  21. Wertz, I. E. et al. Human De-etiolated-1 regulates c-Jun by assembling a CUL4A ubiquitin ligase. Science 303, 1371–1374 (2004)

    Article  ADS  CAS  Google Scholar 

  22. Dornan, D., Shimizu, H., Perkins, N. D. & Hupp, T. R. DNA-dependent acetylation of p53 by the transcription coactivator p300. J. Biol. Chem. 278, 13431–13441 (2003)

    Article  CAS  Google Scholar 

  23. Dornan, D. & Hupp, T. R. Inhibition of p53-dependent transcription by BOX-I phospho-peptide mimetics that bind to p300. EMBO Rep. 2, 139–144 (2001)

    Article  CAS  Google Scholar 

  24. Shimizu, H. et al. The conformationally flexible S9–S10 linker region in the core domain of p53 contains a novel MDM2 binding site whose mutation increases ubiquitination of p53 in vivo. J. Biol. Chem. 277, 28446–28458 (2002)

    Article  CAS  Google Scholar 

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Acknowledgements

We would like to thank T. Hupp for his contribution of reagents and thought-provoking discussions, G. Lozano for p53-/-/MDM2-/- MEFs, S. Benchimol for the Pirh2 antibody and cDNA, J. Chen for H1299 cells, B. Henzel for mass spectrometry support, C. Reed for the generation of COP1 monoclonal antibodies, M. Vasser and P. Ng for oligonucleotide synthesis and purification, A. Waugh for Bioinformatics support, Genentech Protein Engineering and core DNA sequencing facility for support services, and P. Polakis and members of the Dixit lab for advice and encouragement. I.W. is supported by a PSTP fellowship from the University of California, Davis.

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Authors and Affiliations

  1. Department of Molecular Oncology, Genentech, Inc., South San Francisco, 1 DNA Way, California, 94080, USA

    David Dornan, Ingrid Wertz, Harumi Shimizu, Karen O' Rourke & Vishva M. Dixit

  2. Department of Protein Chemistry, Genentech, Inc., 1 DNA Way, South San Francisco, California, 94080, USA

    David Arnott

  3. Department of Pathology, Genentech, Inc., South San Francisco, 1 DNA Way, California, 94080, USA

    Gretchen D. Frantz & Hartmut Koeppen

  4. Department of Molecular Biology, Genentech, Inc., South San Francisco, 1 DNA Way, California, 94080, USA

    Patrick Dowd

  5. Department of Human Physiology, University of California, Davis, California, 95616, USA

    Ingrid Wertz

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  1. David Dornan
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Correspondence to Vishva M. Dixit.

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Supplementary Figures

Supplementary Figure S1: COP1 maintains the ability to degrade p53 in the absence of MDM2 or Pirh2; Supplementary Figure S2: COP1 negative regulation of p53 transactivation activity; Supplementary Figure S3: Ablation of COP1 by two further distinct siRNA oligos causes an accumulation of p53 at the protein level. (PDF 303 kb)

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Dornan, D., Wertz, I., Shimizu, H. et al. The ubiquitin ligase COP1 is a critical negative regulator of p53. Nature 429, 86–92 (2004). https://doi.org/10.1038/nature02514

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