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Trim39 regulates neuronal apoptosis by acting as a SUMO-targeted E3 ubiquitin-ligase for the transcription factor NFATc3

Abstract

NFATc3 is the predominant member of the NFAT family of transcription factors in neurons, where it plays a pro-apoptotic role. Mechanisms controlling NFAT protein stability are poorly understood. Here we identify Trim39 as an E3 ubiquitin-ligase of NFATc3. Indeed, Trim39 binds and ubiquitinates NFATc3 in vitro and in cells where it reduces NFATc3 protein level and transcriptional activity. In contrast, silencing of endogenous Trim39 decreases NFATc3 ubiquitination and increases its activity, thereby resulting in enhanced neuronal apoptosis. We also show that Trim17 inhibits Trim39-mediated ubiquitination of NFATc3 by reducing both the E3 ubiquitin-ligase activity of Trim39 and the NFATc3/Trim39 interaction. Moreover, we identify Trim39 as a new SUMO-targeted E3 ubiquitin-ligase (STUbL). Indeed, mutation of SUMOylation sites in NFATc3 or SUMO-interacting motifs in Trim39 reduces NFATc3/Trim39 interaction and Trim39-induced ubiquitination of NFATc3. In addition, Trim39 preferentially ubiquitinates SUMOylated forms of NFATc3 in vitro. As a consequence, a SUMOylation-deficient mutant of NFATc3 exhibits increased stability and pro-apoptotic activity in neurons. Taken together, these data indicate that Trim39 modulates neuronal apoptosis by acting as a STUbL for NFATc3.

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Fig. 1: Trim39 interacts with both Trim17 and NFATc3.
Fig. 2: Trim39 is an E3 ubiquitin-ligase of NFATc3.
Fig. 3: Trim39 mediates NFATc3 degradation.
Fig. 4: Trim17 inhibits TRIM39-mediated ubiquitination of NFATc3.
Fig. 5: Trim17 reduces the interaction between endogenous Trim39 and NFATc3.
Fig. 6: SUMOylation of NFATc3 favors its ubiquitination and stability.
Fig. 7: Trim39 is a SUMO-targeted E3 ubiquitin-ligase for NFATc3.
Fig. 8: SUMOylation and Trim39 attenuate NFATc3 pro-apoptotic effect in neurons.

Data availability

The data used during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

This article is based upon work from COST Action (PROTEOSTASIS BM1307), supported by COST (European Cooperation in Science and Technology). We would like to thank the staff of the Montpellier Genomic Collection platform for providing human TRIM39 and human TRIM17 cDNA clones. We acknowledge the imaging facility MRI (Montpellier Ressources Imagerie), member of the national infrastructure France-BioImaging infrastructure supported by the French National Research Agency (ANR-10-INBS-04, “Investments for the future”). We are grateful to Frédérique Brockly for the production and purification of recombinant proteins and Dr Olivier Coux for providing ubiquitin mutants. We thank Drs Dimitris Liakopoulos and Manuel Rodriguez for interesting discussions.

Funding

This work was supported by the Centre National de la Recherche Scientifique (CNRS), the Institut National de la Santé et de la Recherche Médicale (INSERM), the Université de Montpellier, La Fondation de l’Association pour la Recherche contre le Cancer (ARC, grant number PJA 20141201882 to SD), La Ligue contre le Cancer (grant number TDUM13665 to BM) and La Fondation pour la Recherche Médicale (grant number FDT201904008340 to MBS).

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MBS, BM and IL designed and performed experiments, and analyzed the data; SM performed experiments; SD and BM conceived the study; SD performed the statistical analysis, wrote and revised the manuscript, and prepared the figures; IR cloned Trim39 and reviewed the manuscript; GB provided material support and contributed in the design of experiments.

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Correspondence to Solange Desagher.

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Basu-Shrivastava, M., Mojsa, B., Mora, S. et al. Trim39 regulates neuronal apoptosis by acting as a SUMO-targeted E3 ubiquitin-ligase for the transcription factor NFATc3. Cell Death Differ 29, 2107–2122 (2022). https://doi.org/10.1038/s41418-022-01002-2

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