title: Shigella subverts the host recycling compartment to rupture its vacuole.
abstract: Shigella enters epithlial cells via internalization into a vacuole. Subsequent vacuolar membrane rupture allows bacterial escape into the cytosol for replication and cell-to-cell spread. Bacterial effectors such as IpgD, a PI(4,5)P2 phosphatase that generates PI(5)P and alters host actin, facilitate this internalization. Here, we identify host proteins involved in Shigella uptake and vacuolar membrane rupture by high-content siRNA screening and subsequently focus on Rab11, a constituent of the recycling compartment. Rab11-positive vesicles are recruited to the invasion site before vacuolar rupture, and Rab11 knockdown dramatically decreases vacuolar membrane rupture. Additionally, Rab11 recruitment is absent and vacuolar rupture is delayed in the ipgD mutant that does not dephosphorylate PI(4,5)P₂ into PI(5)P. Ultrastructural analyses of Rab11-positive vesicles further reveal that ipgD mutant-containing vacuoles become confined in actin structures that likely contribute to delayed vacular rupture. These findings provide insight into the underlying molecular mechanism of vacuole progression and rupture during Shigella invasion.
authors.affiliation: Institut Pasteur, Dynamics of Host-Pathogen interactions Unit, 25 Rue du Dr. Roux, 75724 Paris, France.
authors.forename: Nora
authors.initials: N
authors.lastname: Mellouk
authors.affiliation: Institut Pasteur, Dynamics of Host-Pathogen interactions Unit, 25 Rue du Dr. Roux, 75724 Paris, France.
authors.forename: Allon
authors.initials: A
authors.lastname: Weiner
authors.affiliation: Institut Pasteur, Imagopole, 28 Rue du Dr. Roux, 75724 Paris, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Aulner
authors.affiliation: Institut Pasteur, Imagopole, 28 Rue du Dr. Roux, 75724 Paris, France.
authors.forename: Christine
authors.initials: C
authors.lastname: Schmitt
authors.affiliation: Department of Materials and Interfaces, Weizmann Institute of Sciences, 234 Herzl Street, Rehovot 76100, Israel.
authors.forename: Michael
authors.initials: M
authors.lastname: Elbaum
authors.affiliation: Institut Pasteur, Imagopole, 28 Rue du Dr. Roux, 75724 Paris, France.
authors.forename: Spencer L
authors.initials: SL
authors.lastname: Shorte
authors.affiliation: Institut Pasteur, Imagopole, 28 Rue du Dr. Roux, 75724 Paris, France. Electronic address: anne.danckaert@pasteur.fr.
authors.forename: Anne
authors.initials: A
authors.lastname: Danckaert
authors.affiliation: Institut Pasteur, Dynamics of Host-Pathogen interactions Unit, 25 Rue du Dr. Roux, 75724 Paris, France. Electronic address: jost.enninga@pasteur.fr.
authors.forename: Jost
authors.initials: J
authors.lastname: Enninga
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D003593:Cytoplasm / Q000382:microbiology*; D004705:Endocytosis; D004847:Epithelial Cells / Q000382:microbiology* / Q000502:physiology; D055786:Gene Knockout Techniques; D054884:Host-Pathogen Interactions; D007425:Intracellular Membranes / Q000378:metabolism; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics / Q000378:metabolism*; D012760:Shigella / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D014617:Vacuoles / Q000382:microbiology*; D037521:Virulence Factors / Q000235:genetics / Q000378:metabolism; D020691:rab GTP-Binding Proteins / Q000378:metabolism*; D000099193:rab11 GTP-Binding Proteins
chemical_list: D001426:Bacterial Proteins; D010744:Phosphoric Monoester Hydrolases; D037521:Virulence Factors; D020691:rab GTP-Binding Proteins; D000099193:rab11 GTP-Binding Proteins; C425774:IpgD protein, Shigella flexneri