title: Listeria monocytogenes internalin B activates junctional endocytosis to accelerate intestinal invasion.
abstract: Listeria monocytogenes (Lm) uses InlA to invade the tips of the intestinal villi, a location at which cell extrusion generates a transient defect in epithelial polarity that exposes the receptor for InlA, E-cadherin, on the cell surface. As the dying cell is removed from the epithelium, the surrounding cells reorganize to form a multicellular junction (MCJ) that Lm exploits to find its basolateral receptor and invade. By examining individual infected villi using 3D-confocal imaging, we uncovered a novel role for the second major invasin, InlB, during invasion of the intestine. We infected mice intragastrically with isogenic strains of Lm that express or lack InlB and that have a modified InlA capable of binding murine E-cadherin and found that Lm lacking InlB invade the same number of villi but have decreased numbers of bacteria within each infected villus tip. We studied the mechanism of InlB action at the MCJs of polarized MDCK monolayers and find that InlB does not act as an adhesin, but instead accelerates bacterial internalization after attachment. InlB locally activates its receptor, c-Met, and increases endocytosis of junctional components, including E-cadherin. We show that MCJs are naturally more endocytic than other sites of the apical membrane, that endocytosis and Lm invasion of MCJs depends on functional dynamin, and that c-Met activation by soluble InlB or hepatocyte growth factor (HGF) increases MCJ endocytosis. Also, in vivo, InlB applied through the intestinal lumen increases endocytosis at the villus tips. Our findings demonstrate a two-step mechanism of synergy between Lm's invasins: InlA provides the specificity of Lm adhesion to MCJs at the villus tips and InlB locally activates c-Met to accelerate junctional endocytosis and bacterial invasion of the intestine.
authors.affiliation: Department of Microbiology and Immunology, Stanford University, Stanford, California, United States of America.
authors.forename: Mickey
authors.initials: M
authors.lastname: Pentecost
authors.forename: Jyothi
authors.initials: J
authors.lastname: Kumaran
authors.forename: Partho
authors.initials: P
authors.lastname: Ghosh
authors.forename: Manuel R
authors.initials: MR
authors.lastname: Amieva
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism*; D015820:Cadherins / Q000378:metabolism; D002460:Cell Line; D016764:Cell Polarity / Q000502:physiology; D004285:Dogs; D034281:Dynamins / Q000378:metabolism; D004705:Endocytosis / Q000502:physiology*; D004847:Epithelial Cells / Q000378:metabolism / Q000382:microbiology*; D005260:Female; D007365:Intercellular Junctions / Q000378:metabolism / Q000382:microbiology; D007668:Kidney / Q000166:cytology; D008089:Listeria monocytogenes / Q000378:metabolism*; D008088:Listeriosis / Q000382:microbiology*; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D008871:Microvilli / Q000378:metabolism / Q000382:microbiology; D020794:Receptor Protein-Tyrosine Kinases / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D015820:Cadherins; D008565:Membrane Proteins; C095768:inlB protein, Listeria monocytogenes; C080835:RON protein; D020794:Receptor Protein-Tyrosine Kinases; D034281:Dynamins