title: Role of VAMP3 and VAMP7 in the commitment of Yersinia pseudotuberculosis to LC3-associated pathways involving single- or double-membrane vacuoles.
abstract: Yersinia pseudotuberculosis can replicate inside macrophages by hijacking autophagy and blocking autophagosome acidification. In bone marrow-derived macrophages, the bacteria are mainly observed inside double-membrane vacuoles positive for LC3, a hallmark of autophagy. Here, we address the question of the membrane traffic during internalization of Yersinia investigating the role of vesicle- associated membrane proteins (VAMPs). First, we show that as in epithelial cells, Yersinia pseudotuberculosis replicates mainly in nonacidic LC3-positive vacuoles. Second, in these cells, we unexpectedly found that VAMP3 localizes preferentially to Yersinia-containing vacuoles (YCVs) with single membranes using correlative light-electron microscopy. Third, we reveal the precise kinetics of VAMP3 and VAMP7 association with YCVs positive for LC3. Fourth, we show that VAMP7 knockdown alters LC3's association with single-and multimembrane-YCVs. Finally, in uninfected epithelial cells stimulated for autophagy, VAMP3 overexpression and knockdown led respectively to a lower and higher number of double-membrane, LC3-positive vesicles. Hence, our results highlight the role that VAMPs play in selection of the pathways leading to generation of ultrastructurally different LC3 compartments and pave the way for determining the full set of docking and fusion proteins involved in Yersinia pseudotuberculosis' intravesicular life cycle.
authors.affiliation: Cellular Microbiology of Infectious Pathogens Group; Center for Infection and Immunity of Lille; Institut Pasteur de Lille; Lille, France; CNRS UMR8204; Lille, France; INSERM U1019; Lille, France; University of Lille-Nord de France; Lille, France.
authors.forename: Laure-Anne
authors.initials: LA
authors.lastname: Ligeon
authors.affiliation: Cellular Microbiology of Infectious Pathogens Group; Center for Infection and Immunity of Lille; Institut Pasteur de Lille; Lille, France; CNRS UMR8204; Lille, France; INSERM U1019; Lille, France; University of Lille-Nord de France; Lille, France.
authors.forename: Kevin
authors.initials: K
authors.lastname: Moreau
authors.affiliation: INSERM U1019; Lille, France; BioImaging Center Lille-Nord de France; IFR142; Institut Pasteur de Lille; Lille, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Barois
authors.affiliation: Cellular Microbiology of Infectious Pathogens Group; Center for Infection and Immunity of Lille; Institut Pasteur de Lille; Lille, France; BioImaging Center Lille-Nord de France; IFR142; Institut Pasteur de Lille; Lille, France.
authors.forename: Antonino
authors.initials: A
authors.lastname: Bongiovanni
authors.affiliation: University of Lille-Nord de France; Lille, France; BioImaging Center Lille-Nord de France; IFR142; Institut Pasteur de Lille; Lille, France.
authors.forename: Delphine-Armelle
authors.initials: DA
authors.lastname: Lacorre
authors.affiliation: BioImaging Center Lille-Nord de France; IFR142; Institut Pasteur de Lille; Lille, France; CNRS UM8161; Lille, France.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Werkmeister
authors.affiliation: Institut Jacques Monod; UMR 7592; CNRS; University of Paris Diderot; Paris, France; INSERM ERL U950; Membrane Traffic in Neuronal & Epithelial Morphogenesis Group; Paris, France.
authors.forename: Véronique
authors.initials: V
authors.lastname: Proux-Gillardeaux
authors.affiliation: Institut Jacques Monod; UMR 7592; CNRS; University of Paris Diderot; Paris, France; INSERM ERL U950; Membrane Traffic in Neuronal & Epithelial Morphogenesis Group; Paris, France.
authors.forename: Thierry
authors.initials: T
authors.lastname: Galli
authors.affiliation: Cellular Microbiology of Infectious Pathogens Group; Center for Infection and Immunity of Lille; Institut Pasteur de Lille; Lille, France; CNRS UMR8204; Lille, France; INSERM U1019; Lille, France; University of Lille-Nord de France; Lille, France; BioImaging Center Lille-Nord de France; IFR142; Institut Pasteur de Lille; Lille, France.
authors.forename: Frank
authors.initials: F
authors.lastname: Lafont
mesh_terms: D001343:Autophagy / Q000502:physiology*; D002460:Cell Line; D006801:Humans; D008264:Macrophages / Q000166:cytology; D008854:Microscopy, Electron; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D010588:Phagosomes / Q000648:ultrastructure; D050683:R-SNARE Proteins / Q000378:metabolism*; D015398:Signal Transduction; D014617:Vacuoles / Q000378:metabolism*; D051040:Vesicle-Associated Membrane Protein 3 / Q000378:metabolism*; D015011:Yersinia pseudotuberculosis / Q000378:metabolism*
chemical_list: C477012:MAP1LC3A protein, human; D008869:Microtubule-Associated Proteins; D050683:R-SNARE Proteins; C494946:VAMP3 protein, human; C121241:VAMP7 protein, human; D051040:Vesicle-Associated Membrane Protein 3
keywords: LC3-associated phagosome; SNARE; VAMP3; VAMP7; Yersinia pseudotuberculosis; autophagy; correlative light-electron microscopy