Vinueza-Gavilanes, RodrigoBravo-González, Jorge JuanBasurco, LeyreBoncristiani, ChiaraFernández Irigoyen, JoaquínSantamaría Martínez, EnriqueMarcilla, IrenePérez Mediavilla, AlbertoLuquin, María RosarioVales, AfricaGonzález-Aseguinolaza, GloriaAymerich, María SoledadAragón, TomásArrasate, Montserrat2023-10-162023-10-162023Vinueza-Gavilanes, R., Bravo-González, J. J., Basurco, L., Boncristiani, C., Fernández-Irigoyen, J., Santamaría, E., Marcilla, I., Pérez-Mediavilla, A., Luquin, M. R., Vales, A., González-Aseguinolaza, G., Aymerich, M. S., Aragón, T., Arrasate, M. (2023) Stabilization of 14-3-3 protein-protein interactions with Fusicoccin-A decreases alpha-synuclein dependent cell-autonomous death in neuronal and mouse models. Neurobiology of Disease, 183, 1-15. https://doi.org/10.1016/j.nbd.2023.106166.0969-996110.1016/j.nbd.2023.106166https://academica-e.unavarra.es/handle/2454/46530Synucleinopathies are a group of neurodegenerative diseases without effective treatment characterized by the abnormal aggregation of alpha-synuclein (aSyn) protein. Changes in levels or in the amino acid sequence of aSyn (by duplication/triplication of the aSyn gene or point mutations in the encoding region) cause familial cases of synucleinopathies. However, the specific molecular mechanisms of aSyn-dependent toxicity remain unclear. Increased aSyn protein levels or pathological mutations may favor abnormal protein-protein interactions (PPIs) that could either promote neuronal death or belong to a coping response program against neurotoxicity. Therefore, the identification and modulation of aSyn-dependent PPIs can provide new therapeutic targets for these diseases. To identify aSyn-dependent PPIs we performed a proximity biotinylation assay based on the promiscuous biotinylase BioID2. When expressed as a fusion protein, BioID2 biotinylates by proximity stable and transient interacting partners, allowing their identification by streptavidin affinity purification and mass spectrometry. The aSyn interactome was analyzed using BioID2-tagged wild-type (WT) and pathological mutant E46K aSyn versions in HEK293 cells. We found the 14-3-3 epsilon isoform as a common protein interactor for WT and E46K aSyn. 14‐3-3 epsilon correlates with aSyn protein levels in brain regions of a transgenic mouse model overexpressing WT human aSyn. Using a neuronal model in which aSyn cell-autonomous toxicity is quantitatively scored by longitudinal survival analysis, we found that stabilization of 14‐3-3 protein-proteins interactions with Fusicoccin-A (FC-A) decreases aSyn-dependent toxicity. Furthermore, FC-A treatment protects dopaminergic neuronal somas in the substantia nigra of a Parkinson's disease mouse model. Based on these results, we propose that the stabilization of 14‐3-3 epsilon interaction with aSyn might reduce aSyn toxicity, and highlight FC-A as a potential therapeutic compound for synucleinopathies.application/pdfeng© 2023 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY-NC-ND license.14-3-3 epsilonAdenoassociated virus AAV9Alpha-synucleinBioID2Fusicoccin-ALongitudinal survival analysisNeuronal deathParkinson's diseaseProximity biotinylationSynucleinopathiesinfo:eu-repo/semantics/article2023-10-16info:eu-repo/semantics/openAccess