title: Proteomic analysis of the Simkania-containing vacuole: the central role of retrograde transport.
abstract: Simkania negevensis is an obligate intracellular bacterial pathogen that grows in amoeba or human cells within a membrane-bound vacuole forming endoplasmic reticulum (ER) contact sites. The membrane of this Simkania-containing vacuole (SnCV) is a critical host-pathogen interface whose origin and molecular interactions with cellular organelles remain poorly defined. We performed proteomic analysis of purified ER-SnCV-membranes using label free LC-MS(2) to define the pathogen-containing organelle composition. Of the 1,178 proteins of human and 302 proteins of Simkania origin identified by this strategy, 51 host cell proteins were enriched or depleted by infection and 57 proteins were associated with host endosomal transport pathways. Chemical inhibitors that selectively interfere with trafficking at the early endosome-to-trans-Golgi network (TGN) interface (retrograde transport) affected SnCV formation, morphology and lipid transport. Our data demonstrate that Simkania exploits early endosome-to-TGN transport for nutrient acquisition and growth.
authors.affiliation: Department of Microbiology, University of Würzburg, Biocenter, Am Hubland, D-97074, Würzburg, Germany.
authors.forename: Jo-Ana
authors.initials: JA
authors.lastname: Herweg
authors.affiliation: DSV, iBiTec-S, LabEx LERMIT, CEA, F-91191, Gif sur Yvette, France.
authors.forename: Valérie
authors.initials: V
authors.lastname: Pons
authors.affiliation: Department of Microbial Proteomics, University of Greifswald, Institute of Microbiology, Friedrich-Ludwig-Jahn-Straße 15, D-17487, Greifswald, Germany.
authors.forename: Dörte
authors.initials: D
authors.lastname: Becher
authors.affiliation: Microbial Physiology/Molecular Biology, University of Greifswald, Institute of Microbiology, Friedrich-Ludwig-Jahn-Straße 15, D-17487, Greifswald, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Hecker
authors.affiliation: Division of Electron Microscopy, University of Würzburg, Biocenter, Am Hubland, D-97074, Würzburg, Germany.
authors.forename: Georg
authors.initials: G
authors.lastname: Krohne
authors.affiliation: DSV, iBiTec-S, LabEx LERMIT, CEA, F-91191, Gif sur Yvette, France.
authors.forename: Julien
authors.initials: J
authors.lastname: Barbier
authors.affiliation: Department of Molecular Biology, Max-Planck-Institute for Infection Biology, Charitéplatz 1, D-10117, Berlin, Germany.
authors.forename: Hilmar
authors.initials: H
authors.lastname: Berger
authors.affiliation: Department of Microbiology, University of Würzburg, Biocenter, Am Hubland, D-97074, Würzburg, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Rudel
authors.affiliation: Department of Microbiology, University of Würzburg, Biocenter, Am Hubland, D-97074, Würzburg, Germany.|Department of Molecular Biology, Max-Planck-Institute for Infection Biology, Charitéplatz 1, D-10117, Berlin, Germany.
authors.forename: Adrian
authors.initials: A
authors.lastname: Mehlitz
mesh_terms: D002695:Chlamydiales / Q000254:growth & development*; D002853:Chromatography, Liquid; D006367:HeLa Cells; D006801:Humans; D007425:Intracellular Membranes / Q000737:chemistry*; D013058:Mass Spectrometry; D020543:Proteome / Q000032:analysis*; D040901:Proteomics; D014617:Vacuoles / Q000737:chemistry* / Q000382:microbiology*
chemical_list: D020543:Proteome