title: Listeria monocytogenes triggers the cell surface expression of Gp96 protein and interacts with its N terminus to support cellular infection.
abstract: Listeria monocytogenes is an intracellular food-borne pathogen causing listeriosis in humans. This bacterium deploys an arsenal of virulence factors that act in concert to promote cellular infection. Bacterial surface proteins are of primary importance in the process of host cell invasion. They interact with host cellular receptors, inducing/modulating specific cellular responses. We previously identified Vip, a Listeria surface protein covalently attached to the bacterial cell wall acting as a key virulence factor. We have shown that Vip interacts with Gp96 localized at the surface of host cells during invasion and that this interaction is critical for a successful infection in vivo. To better understand the importance of Vip-Gp96 interaction during infection, we aimed to characterize this interaction at the molecular level. Here we demonstrate that, during infection, L. monocytogenes triggers the cellular redistribution of Gp96, inducing its exposure at the cell surface. Upon infection, Gp96 N-terminal domain is exposed to the extracellular milieu in L2071 fibroblasts and interacts with Vip expressed by Listeria. We identified Gp96 (Asp(1)-Leu(170)) as sufficient to interact with Vip; however, we also showed that the region Tyr(179)-Leu(390) of Gp96 is important for the interaction. Our findings unravel the Listeria-induced surface expression of Gp96 and the topology of its insertion on the plasma membrane and improve our knowledge on the Vip-Gp96 interaction during Listeria infection.
authors.affiliation: Group of Molecular Microbiology, Instituto de Biologia Molecular e Celular, Universidade do Porto, 4150-180 Porto, Portugal.
authors.forename: Mariana
authors.initials: M
authors.lastname: Martins
authors.forename: Rafael
authors.initials: R
authors.lastname: Custódio
authors.forename: Ana
authors.initials: A
authors.lastname: Camejo
authors.forename: Maria Teresa
authors.initials: MT
authors.lastname: Almeida
authors.forename: Didier
authors.initials: D
authors.lastname: Cabanes
authors.forename: Sandra
authors.initials: S
authors.lastname: Sousa
mesh_terms: D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D001426:Bacterial Proteins / Q000378:metabolism; D018938:Caco-2 Cells; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D008089:Listeria monocytogenes / Q000187:drug effects / Q000502:physiology*; D008088:Listeriosis / Q000378:metabolism* / Q000382:microbiology*; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D011485:Protein Binding / Q000187:drug effects; D017434:Protein Structure, Tertiary
chemical_list: D000906:Antibodies; D001426:Bacterial Proteins; D008562:Membrane Glycoproteins; C503594:Vip protein, Listeria monocytogenes; C053387:endoplasmin