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The RanBP2/RanGAP1-SUMO complex gates β-arrestin2 nuclear entry to regulate the Mdm2-p53 signaling axis

Abstract

Mdm2 antagonizes the tumor suppressor p53. Targeting the Mdm2-p53 interaction represents an attractive approach for the treatment of cancers with functional p53. Investigating mechanisms underlying Mdm2-p53 regulation is therefore important. The scaffold protein β-arrestin2 (β-arr2) regulates tumor suppressor p53 by counteracting Mdm2. β-arr2 nucleocytoplasmic shuttling displaces Mdm2 from the nucleus to the cytoplasm resulting in enhanced p53 signaling. β-arr2 is constitutively exported from the nucleus, via a nuclear export signal, but mechanisms regulating its nuclear entry are not completely elucidated. β-arr2 can be SUMOylated, but no information is available on how SUMO may regulate β-arr2 nucleocytoplasmic shuttling. While we found β-arr2 SUMOylation to be dispensable for nuclear import, we identified a non-covalent interaction between SUMO and β-arr2, via a SUMO interaction motif (SIM), that is required for β-arr2 cytonuclear trafficking. This SIM promotes association of β-arr2 with the multimolecular RanBP2/RanGAP1-SUMO nucleocytoplasmic transport hub that resides on the cytoplasmic filaments of the nuclear pore complex. Depletion of RanBP2/RanGAP1-SUMO levels result in defective β-arr2 nuclear entry. Mutation of the SIM inhibits β-arr2 nuclear import, its ability to delocalize Mdm2 from the nucleus to the cytoplasm and enhanced p53 signaling in lung and breast tumor cell lines. Thus, a β-arr2 SIM nuclear entry checkpoint, coupled with active β-arr2 nuclear export, regulates its cytonuclear trafficking function to control the Mdm2-p53 signaling axis.

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Fig. 1: β-arr2 is SUMOylated in cells.
Fig. 2: β-arr2 is SUMOylated on lysine 295.
Fig. 3: β-arr2 contains a SIM in its N-domain.
Fig. 4: The SIM but not SUMOylation on lysine 295 is required for β-arr2 nuclear entry.
Fig. 5: SUMO1 fusion to β-arr2∆SIM does not rescue nuclear import but an NLS fusion does.
Fig. 6: The β-arr2 SIM enhances association with the RanBP2/RanGAP1-SUMO complex and RanBP2/RanGAP1-SUMO depletion inhibits β-arr2 nuclear entry.
Fig. 7: A functional SIM domain is required for β-arr2-mediated cytoplasmic delocalization of Mdm2.
Fig. 8: The β-arr2 SIM domain is required for increased p53 signaling.

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Acknowledgements

We thank Dr. A. Benmerah for helpful discussion, Dr. J. Liotard for excellent technical assistance, and the Institut Cochin Imaging (IMAG’IC) and Sequencing platforms (GENOM’IC). The Institut Cochin lab is part of the Who am I? laboratory of excellence (grant ANR-11-LABX-0071), funded by the “Investments for the Future” program operated by The French National Research Agency (grant ANR-11-IDEX-0005-01). This work was funded by grants from the Fondation ARC pour la Recherche sur le Cancer (“Projet ARC” to MGHS), Ligue contre le Cancer (to MGHS), Royal Society (“International Joint Project Scheme” to MGHS and GSB), France Canada Research Fund (to MGHS and SA), CNRS, and INSERM. The work in the laboratory of M.B. was supported by a CIHR Foundation (FDN148431) grant. M.B. holds the Canada Research Chair in Signal transduction and Molecular Pharmacology. EBT was funded by MESR and Fondation ARC pour la Recherche sur le Cancer doctoral fellowships.

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EBT, ML, BS, JSP, GSB, HE, MB, SA, SM, and MGHS designed research. EBT, ML, JSP, BS, MK, KS, JF, AP, and MGHS performed research. EBT, ML, BS, JSP, CA, ELF, AZ, LG, GSB, HE, MB, SA, SM, and MGHS analyzed data. MGHS supervised the project. EBT and MGHS wrote the paper, which was subsequently reviewed by all other authors.

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Correspondence to Mark G. H. Scott.

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Blondel-Tepaz, E., Leverve, M., Sokrat, B. et al. The RanBP2/RanGAP1-SUMO complex gates β-arrestin2 nuclear entry to regulate the Mdm2-p53 signaling axis. Oncogene 40, 2243–2257 (2021). https://doi.org/10.1038/s41388-021-01704-w

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