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

Abstract

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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Figure 1: Arsenic induces SUMOylation, ubiquitination and degradation of PML.
Figure 2: Role of SUMOs and PML SUMOylation sites in arsenic-induced degradation.
Figure 3: RNF4 degrades PML on nuclear bodies.
Figure 4: PML–RARα degradation is required for arsenic-induced differentiation ex vivo.

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Acknowledgements

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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Correspondence to Hugues de Thé.

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Lallemand-Breitenbach, V., Jeanne, M., Benhenda, S. et al. Arsenic degrades PML or PML–RARα through a SUMO-triggered RNF4/ubiquitin-mediated pathway. Nat Cell Biol 10, 547–555 (2008). https://doi.org/10.1038/ncb1717

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