Stabilization of 14-3-3 protein-protein interactions with Fusicoccin-A decreases alpha-synuclein dependent cell-autonomous death in neuronal and mouse models

Rodrigo Vinueza-Gavilanes; Jorge Juan Bravo-González; Leyre Basurco; Chiara Boncristiani; Joaquín Fernández-Irigoyen; Enrique Santamaría; Irene Marcilla; Alberto Pérez-Mediavilla; María Rosario Luquin; Africa Vales; Gloria González-Aseguinolaza; María Soledad Aymerich; Tomás Aragón; Montserrat Arrasate

doi:10.1016/j.nbd.2023.106166

Stabilization of 14-3-3 protein-protein interactions with Fusicoccin-A decreases alpha-synuclein dependent cell-autonomous death in neuronal and mouse models

Neurobiol Dis. 2023 Jul:183:106166. doi: 10.1016/j.nbd.2023.106166. Epub 2023 May 26.

Authors

Affiliations

¹ Gene Therapy and Regulation of Gene Expression Program, Center for Applied Medical Research (CIMA), University of Navarra, Pamplona, Spain. Electronic address: [email protected].
² Gene Therapy and Regulation of Gene Expression Program, Center for Applied Medical Research (CIMA), University of Navarra, Pamplona, Spain. Electronic address: [email protected].
³ Gene Therapy and Regulation of Gene Expression Program, Center for Applied Medical Research (CIMA), University of Navarra, Pamplona, Spain; Biochemistry and Genetics Department, School of Sciences, University of Navarra, Pamplona, Spain. Electronic address: [email protected].
⁴ Department of Life Sciences, Università degli Studi di Trieste, Trieste, Italy. Electronic address: [email protected].
⁵ Proteored-Institute of Health Carlos III (ISCIII), Clinical Neuroproteomics Unit, Navarrabiomed, Navarra Health Department, Public University of Navarra, Pamplona, Spain; IdiSNA (Navarra Institute for Health Research), Pamplona, Spain. Electronic address: [email protected].
⁶ Proteored-Institute of Health Carlos III (ISCIII), Clinical Neuroproteomics Unit, Navarrabiomed, Navarra Health Department, Public University of Navarra, Pamplona, Spain; IdiSNA (Navarra Institute for Health Research), Pamplona, Spain. Electronic address: [email protected].
⁷ Gene Therapy and Regulation of Gene Expression Program, Center for Applied Medical Research (CIMA), University of Navarra, Pamplona, Spain; IdiSNA (Navarra Institute for Health Research), Pamplona, Spain. Electronic address: [email protected].
⁸ Gene Therapy and Regulation of Gene Expression Program, Center for Applied Medical Research (CIMA), University of Navarra, Pamplona, Spain; Biochemistry and Genetics Department, School of Sciences, University of Navarra, Pamplona, Spain; IdiSNA (Navarra Institute for Health Research), Pamplona, Spain. Electronic address: [email protected].
⁹ Department of Neurology, Clínica Universidad de Navarra, University of Navarra, Pamplona, Spain; IdiSNA (Navarra Institute for Health Research), Pamplona, Spain. Electronic address: [email protected].
¹⁰ Gene Therapy and Regulation of Gene Expression Program, Center for Applied Medical Research (CIMA), University of Navarra, Pamplona, Spain; IdiSNA (Navarra Institute for Health Research), Pamplona, Spain. Electronic address: [email protected].
¹¹ Gene Therapy and Regulation of Gene Expression Program, Center for Applied Medical Research (CIMA), University of Navarra, Pamplona, Spain; IdiSNA (Navarra Institute for Health Research), Pamplona, Spain. Electronic address: [email protected].
¹² Gene Therapy and Regulation of Gene Expression Program, Center for Applied Medical Research (CIMA), University of Navarra, Pamplona, Spain; Biochemistry and Genetics Department, School of Sciences, University of Navarra, Pamplona, Spain; IdiSNA (Navarra Institute for Health Research), Pamplona, Spain. Electronic address: [email protected].
¹³ Gene Therapy and Regulation of Gene Expression Program, Center for Applied Medical Research (CIMA), University of Navarra, Pamplona, Spain; IdiSNA (Navarra Institute for Health Research), Pamplona, Spain. Electronic address: [email protected].
¹⁴ Gene Therapy and Regulation of Gene Expression Program, Center for Applied Medical Research (CIMA), University of Navarra, Pamplona, Spain; Department of Pathology, Anatomy and Physiology, School of Medicine, University of Navarra, Pamplona, Spain; IdiSNA (Navarra Institute for Health Research), Pamplona, Spain. Electronic address: [email protected].

PMID: 37245833
DOI: 10.1016/j.nbd.2023.106166

Abstract

Synucleinopathies 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.

Keywords: 14-3-3 epsilon; Adenoassociated virus AAV9; Alpha-synuclein; BioID2; Fusicoccin-A; Longitudinal survival analysis; Neuronal death; Parkinson's disease; Proximity biotinylation; Synucleinopathies.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

14-3-3 Proteins / genetics
14-3-3 Proteins / metabolism
Animals
Dopaminergic Neurons / metabolism
HEK293 Cells
Humans
Mice
Mice, Transgenic
Synucleinopathies*
alpha-Synuclein* / metabolism

Substances

alpha-Synuclein
14-3-3 Proteins
fusicoccin