Abstract
Oleate, the most abundant endogenous and dietary cis-unsaturated fatty acid, has the atypical property to cause the redistribution of microtubule-associated proteins 1A/1B light chain 3B (referred to as LC3) to the trans-Golgi network (TGN), as shown here. A genome-wide screen identified multiple, mostly Golgi transport-related genes specifically involved in the oleate-induced relocation of LC3 to the Golgi apparatus. Follow-up analyses revealed that oleate also caused the retention of secreted proteins in the TGN, as determined in two assays in which the secretion of proteins was synchronized, (i) an assay involving a thermosensitive vesicular stomatitis virus G (VSVG) protein that is retained in the endoplasmic reticulum (ER) until the temperature is lowered, and (ii) an isothermic assay involving the reversible retention of the protein of interest in the ER lumen and that was used both in vitro and in vivo. A pharmacological screen searching for agents that induce LC3 aggregation at the Golgi apparatus led to the identification of “oleate mimetics” that share the capacity to block conventional protein secretion. In conclusion, oleate represents a class of molecules that act on the Golgi apparatus to cause the recruitment of LC3 and to stall protein secretion.
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Acknowledgements
GC is supported by a scholarship of the Fondation pour la Recherche Médicale (FRM FDT202001011060). GK is supported by the Ligue contre le Cancer (équipe labellisée); Agence National de la Recherche (ANR)—Projets blancs; ANR under the frame of E-Rare-2, the ERA-Net for Research on Rare Diseases; Association pour la recherche sur le cancer (ARC); Cancéropôle Ile-de-France; Chancellerie des universités de Paris (Legs Poix), Fondation pour la Recherche Médicale (FRM); a donation by Elior; European Research Area Network on Cardiovascular Diseases (ERA-CVD, MINOTAUR); Gustave Roussy Odyssea, the European Union Horizon 2020 Project Oncobiome; Fondation Carrefour; High-end Foreign Expert Program in China (GDW20171100085 and GDW20181100051), Institut National du Cancer (INCa); Inserm (HTE); Institut Universitaire de France; LeDucq Foundation; the LabEx Immuno-Oncology; the RHU Torino Lumière; the Seerave Foundation; the SIRIC Stratified Oncology Cell DNA Repair and Tumor Immune Elimination (SOCRATE); and the SIRIC Cancer Research and Personalized Medicine (CARPEM).
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Conceptualization, GK, AS, and GC; Methodology, GK, AS, GC, JH, ML, and PL; Investigation, GC, KM, PL, LZ, WX, and SZ; Formal analysis, AS, GC, and ML; Writing—Original Draft, GK, GC, and AS; Writing—Review & Editing, GK, GC, OK, and JH; Funding acquisition, GK; Supervision, OK and GK.
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GK and OK are cofounders of Samsara Therapeutics. GK is a founder of everImmune and Therafast Bio.
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Animal experiments were conducted in compliance with the EU Directive 63/2010/EU from the European Parliament and were approved by the local Ethics Committee of the Gustave Roussy Cancer Campus (CEEA IRCIV/IGR no. 26, registered at the French Ministry of Research.
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Cerrato, G., Leduc, M., Müller, K. et al. Oleate-induced aggregation of LC3 at the trans-Golgi network is linked to a protein trafficking blockade. Cell Death Differ 28, 1733–1752 (2021). https://doi.org/10.1038/s41418-020-00699-3
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DOI: https://doi.org/10.1038/s41418-020-00699-3
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