title: The DUF582 Proteins ofChlamydia trachomatisBind to Components of the ESCRT Machinery, Which Is Dispensable for Bacterial GrowthIn vitro.
abstract: Chlamydiaeare Gram negative bacteria that develop exclusively inside eukaryotic host cells, within a membrane-bounded compartment. Members of the familyChlamydiaceae, such asChlamydia trachomatis, are pathogenic species infecting vertebrates. They have a very reduced genome and exploit the capacities of their host for their own development, mainly through the secretion of proteins tailored to interfere with eukaryotic processes, called effector proteins. AllChlamydiaceaepossess genes coding for four to five effectors that share a domain of unknown function (DUF582). Here we show that four of these effectors, which represent the conserved set in allChlamydiaceae, accumulate in the infectious form ofC. trachomatis, and are therefore likely involved in an early step of the developmental cycle. The fifth member of the family, CT621, is specific toC. trachomatis, and is secreted during the growth phase. Using a two-hybrid screen in yeast we identified an interaction between the host protein Hrs and the DUF582, which we confirmed by co-immunoprecipitations in co-transfected mammalian cells. Furthermore, we provide biochemical evidence that a second domain of one of the DUF582 proteins, CT619, binds the host protein Tsg101. Hrs and Tsg101 are both implicated in a well conserved machinery of the eukaryotic cell called the ESCRT machinery, which is involved in several cellular processes requiring membrane constriction. Using RNA interference targeting proteins implicated at different stages of ESCRT-driven processes, or inhibition by expression of a dominant negative mutant of VPS4, we demonstrated that this machinery was dispensable for bacterial entry, multiplication and differentiation into infectious progeny, and for uptake of glycogen into the parasitophorous vacuole. In light of these observations we discuss how the DUF582 proteins might target the ESCRT machinery during infection.
authors: Institut Pasteur, Unité de Biologie Cellulaire de l'Infection MicrobienneParis, France; CNRS UMR 3691Paris, France; Université Pierre et Marie Curie, Cellule Pasteur UPMCParis, France.
authors: François
authors: F
authors: Vromman
authors: Institut Pasteur, Unité de Biologie Cellulaire de l'Infection MicrobienneParis, France; CNRS UMR 3691Paris, France.
authors: Stéphanie
authors: S
authors: Perrinet
authors: Institut Pasteur, Unité de Biologie Cellulaire de l'Infection MicrobienneParis, France; CNRS UMR 3691Paris, France.
authors: Lena
authors: L
authors: Gehre
authors: Institut Pasteur, Unité de Biologie Cellulaire de l'Infection MicrobienneParis, France; CNRS UMR 3691Paris, France.
authors: Agathe
authors: A
authors: Subtil
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D002692:Chlamydia trachomatis / Q000254:growth & development*; D017124:Conserved Sequence; D004268:DNA-Binding Proteins / Q000378:metabolism; D056827:Endosomal Sorting Complexes Required for Transport / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D054884:Host-Pathogen Interactions; D006801:Humans; D047468:Immunoprecipitation; D010750:Phosphoproteins / Q000378:metabolism*; D011485:Protein Binding; D000072417:Protein Domains; D025941:Protein Interaction Mapping; D014157:Transcription Factors / Q000378:metabolism; D020798:Two-Hybrid System Techniques; D037521:Virulence Factors / Q000235:genetics / Q000378:metabolism*; D000099177:Tsg101 Protein
chemical_list: D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D056827:Endosomal Sorting Complexes Required for Transport; D010750:Phosphoproteins; D014157:Transcription Factors; D037521:Virulence Factors; D000099177:Tsg101 Protein; C096164:hepatocyte growth factor-regulated tyrosine kinase substrate
keywords: Chlamydia trachomatis; ESCRT; Hrs; Tsg101; host pathogen interactions; type III effectors