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Arsenic degrades PML or PML–RARα through a SUMO-triggered RNF4/ubiquitin-mediated pathway

Nature Cell Biology volume 10, pages 547555 (2008) | Download Citation



In acute promyelocytic leukaemia (APL), arsenic trioxide induces degradation of the fusion protein encoded by the PML–RARA oncogene, differentiation of leukaemic cells and produces clinical remissions. SUMOylation of its PML moiety was previously implicated, but the nature of the degradation pathway involved and the role of PML–RARα catabolism in the response to therapy have both remained elusive. Here, we demonstrate that arsenic-induced PML SUMOylation triggers its Lys 48-linked polyubiquitination and proteasome-dependent degradation. When exposed to arsenic, SUMOylated PML recruits RNF4, the human orthologue of the yeast SUMO-dependent E3 ubiquitin-ligase, as well as ubiquitin and proteasomes onto PML nuclear bodies. Arsenic-induced differentiation is impaired in cells transformed by a non-degradable PML–RARα SUMOylation mutant or in APL cells transduced with a dominant-negative RNF4, directly implicating PML–RARα catabolism in the therapeutic response. We thus identify PML as the first protein degraded by SUMO-dependent polyubiquitination. As PML SUMOylation recruits not only RNF4, ubiquitin and proteasomes, but also many SUMOylated proteins onto PML nuclear bodies, these domains could physically integrate the SUMOylation, ubiquitination and degradation pathways.

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  1. 1.

    , , & How acute promyelocytic leukaemia revived arsenic. Nature Rev. Cancer 2, 705–713. (2002).

  2. 2.

    et al. Arsenic-induced PML targeting onto nuclear bodies: implications for the treatment of acute promyelocytic leukemia. Proc. Natl Acad. Sci. USA 94, 3978–3983. (1997).

  3. 3.

    , & Conjugation with the ubiquitin-related modifier SUMO-1 regulates the partitioning of PML within the nucleus. EMBO J. 17, 61–70 (1998).

  4. 4.

    et al. Arsenic trioxide as an inducer of apoptosis and loss of PML/RARα protein in acute promyelocytic leukemia cells. J. Natl Cancer Inst. 90, 124–133 (1998).

  5. 5.

    et al. Role of promyelocytic leukemia (PML) SUMOylation in nuclear body formation, 11S proteasome recruitment, and As(2)O(3)-induced PML or PML/retinoic acid receptor α Degradation. J. Exp. Med. 193, 1361–1372. (2001).

  6. 6.

    et al. Use of arsenic trioxide (As2O3) in the treatment of acute promyelocytic leukemia (APL): I. As2O3 exerts dose-dependent dual effects on APL cells. Blood 89, 3345–3353 (1997).

  7. 7.

    , , & , JNK activation is a mediator of arsenic trioxide-induced apoptosis in acute promyelocytic leukemia cells. Blood 103, 3496–3502 (2004).

  8. 8.

    , , , & Mechanisms of action of arsenic trioxide. Cancer Res. 62, 3893–3903 (2002).

  9. 9.

    , , , & PIC1, a novel ubiquitin-like protein which interacts with the PML component of a multiprotein complex that is disrupted in acute promyelocytic leukaemia. Oncogene 13, 971–982 (1996).

  10. 10.

    & Functional heterogeneity of small ubiquitin-related protein modifiers SUMO-1 versus SUMO-2/3. J. Biol. Chem. 275, 6252–6258 (2000).

  11. 11.

    , & Conserved function of RNF4 family proteins in eukaryotes: targeting a ubiquitin ligase to SUMOylated proteins. EMBO J. 26, 4102–4112 (2007).

  12. 12.

    et al. SUMO-targeted ubiquitin ligases in genome stability. EMBO J. 26, 4089–4101 (2007).

  13. 13.

    et al. Ubiquitin-dependent proteolytic control of SUMO conjugates. J. Biol. Chem. 282, 34167–34175 (2007).

  14. 14.

    et al. The yeast Hex3.Slx8 heterodimer is a ubiquitin ligase stimulated by substrate sumoylation. J. Biol. Chem. 282, 34176–34184 (2007).

  15. 15.

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

  16. 16.

    et al. Identification of three major sentrinization sites in PML. J. Biol. Chem. 41, 26675–26682 (1998).

  17. 17.

    , , , & The mechanisms of PML-nuclear body formation. Mol. Cell 24, 331–339 (2006).

  18. 18.

    et al. Intracellular localization of proteasomal degradation of a viral antigen. J. Cell Biol. 146, 113–124 (1999).

  19. 19.

    , , & Interferon γ regulates accumulation of the proteasome activator PA28 and immunoproteasomes at nuclear PML bodies. J. Cell Sci. 114, 29–36 (2001).

  20. 20.

    & Identification of the preferential ubiquitination site and ubiquitin-dependent degradation signal of Rpn4. J. Biol. Chem. 281, 10657–10662 (2006).

  21. 21.

    et al. SUMO-1 modification of the acute promyelocytic leukaemia protein PML: implications for nuclear localisation. J. Cell Sci. 112, 381–393 (1999).

  22. 22.

    et al. Polymeric chains of SUMO-2 and SUMO-3 are conjugated to protein substrates by SAE1/SAE2 and Ubc9. J. Biol. Chem. 276, 35368–35374 (2001).

  23. 23.

    , , , & Transcriptional co-regulator SNURF (RNF4) possesses ubiquitin E3 ligase activity. FEBS Lett. 560, 56–62 (2004).

  24. 24.

    , , , & Specification of SUMO1- and SUMO2-interacting motifs. J. Biol. Chem. 281, 16117–16127 (2006).

  25. 25.

    , , & SUMO-1 promotes association of SNURF (RNF4) with PML nuclear bodies. Exp. Cell Res. 304, 224–233 (2005).

  26. 26.

    et al. A sumoylation site in PML/RAR α is essential for leukemic transformation. Cancer Cell 7, 143–153 (2005).

  27. 27.

    , , & SUMO-specific protease 1 is essential for stabilization of HIF1α during hypoxia. Cell 131, 584–595 (2007).

  28. 28.

    et al. Stabilization of PML nuclear localization by conjugation and oligomerization of SUMO-3. Oncogene 24, 5401–5413 (2005).

  29. 29.

    , , , & Noncovalent interaction between Ubc9 and SUMO promotes SUMO chain formation. EMBO J. 26, 2797–2807 (2007).

  30. 30.

    et al. In vivo identification of human small ubiquitin-like modifier polymerization sites by high accuracy mass spectrometry and an in vitro to in vivo strategy. Mol. Cell Proteomics 7, 132–144 (6 April 2008).

  31. 31.

    et al. RNF4 is a poly-SUMO-specific E3 ubiquitin ligase required for arsenic-induced PML degradation. Nature Cell Biol. advance online publication doi:10.1038/ncb1716 (2008).

  32. 32.

    et al. Clastosome: A subtype of nuclear body enriched in 19S and 20S proteasomes, ubiquitin and protein substrates of proteasome. Mol. Biol. Cell 13, 2771–2782 (2002).

  33. 33.

    , & PML residue lysine 160 is required for the degradation of PML induced by herpes simplex virus type 1 regulatory protein ICP0. J. Virol. 77, 8686–8694 (2003).

  34. 34.

    et al. Automated identification of SUMOylation sites using mass spectrometry and SUMmOn pattern recognition software. Nature Methods 3, 533–539 (2006).

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This work was supported by Inca, Canceropôle Ile de France, Ligue Nationale Contre le Cancer (LNCC, 863 program (2006 AA02Z150) and the National Natural Science Foundation of China (30525006). B. R. was supported by the Canada Research Chairs Program, Canada Fund for Innovation, Ontario Innovation Trust and the Canadian Institutes for Health Research. M. J. has a PhD scholarship from Region Ile de France. V. L.-B. is an INSERM staff scientist. We warmly thank J. Godet (LNCC) for her continuous support of this project. We thank J. Palvimo for providing an expression vector and antibodies for RNF4. We are most grateful to R. Hay for providing one of the RNF4 siRNAs and discussion of unpublished data. The role of P. G. Pedrioli in designing the SUMmOn program is gratefully acknowledged, as is the critical help of Stéphanie Duffort. Some experiments were initiated in the laboratory of Benjamin G. Neel, supported by R37 CA49152, with the help of Ricky Chan. We thank N. Setterblad and the Service Commun D'imagerie Cellulaire et Moléculaire for their help and J. C. Gluckman and F. Sigaux for reading the manuscript.

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Author notes

    • Valérie Lallemand-Breitenbach
    •  & Marion Jeanne

    These authors contributed equally to this work.


  1. Université de Paris 7/CNRS UMR 7151, Equipe Labellisée N°11 Ligue Nationale Contre le Cancer, Hôpital St. Louis, 1, Av. C. Vellefaux 75475 Paris CEDEX 10 France.

    • Valérie Lallemand-Breitenbach
    • , Marion Jeanne
    • , Shirine Benhenda
    • , Rihab Nasr
    • , Ming Lei
    • , Laurent Peres
    • , Jun Zhou
    • , Jun Zhu
    •  & Hugues de Thé
  2. CNRS Laboratoire Associé MPC, Shanghai Institute of Hematology, Rui Jin Hospital, 197 Rui Jin Road, 200025 Shanghai China.

    • Jun Zhou
    • , Jun Zhu
    •  & Hugues de Thé
  3. Ontario Cancer Institute and McLaughlin Centre for Molecular Medicine 101 College St., MaRS TMDT 9-805, Toronto, ON M5G 1L7 Canada.

    • Brian Raught
  4. Institut Universitaire de France.

    • Hugues de Thé


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Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Hugues de Thé.

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