title: Host-Microbe Protein Interactions during Bacterial Infection.
abstract: Interspecies protein-protein interactions are essential mediators of infection. While bacterial proteins required for host cell invasion and infection can be identified through bacterial mutant library screens, information about host target proteins and interspecies complex structures has been more difficult to acquire. Using an unbiased chemical crosslinking/mass spectrometry approach, we identified interspecies protein-protein interactions in human lung epithelial cells infected with Acinetobacter baumannii. These efforts resulted in identification of 3,076 crosslinked peptide pairs and 46 interspecies protein-protein interactions. Most notably, the key A. baumannii virulence factor, OmpA, was identified as crosslinked to host proteins involved in desmosomes, specialized structures that mediate host cell-to-cell adhesion. Co-immunoprecipitation and transposon mutant experiments were used to verify these interactions and demonstrate relevance for host cell invasion and acute murine lung infection. These results shed new light on A. baumannii-host protein interactions and their structural features, and the presented approach is generally applicable to other systems.
authors.affiliation: Department of Genome Sciences, University of Washington School of Medicine, Seattle, WA 98195, USA.
authors.forename: Devin K
authors.initials: DK
authors.lastname: Schweppe
authors.affiliation: Departments of Medicine and Microbiology, University of Washington School of Medicine, Seattle, WA 98195, USA.
authors.forename: Christopher
authors.initials: C
authors.lastname: Harding
authors.affiliation: Department of Genome Sciences, University of Washington School of Medicine, Seattle, WA 98195, USA.
authors.forename: Juan D
authors.initials: JD
authors.lastname: Chavez
authors.affiliation: Department of Genome Sciences, University of Washington School of Medicine, Seattle, WA 98195, USA.
authors.forename: Xia
authors.initials: X
authors.lastname: Wu
authors.affiliation: Department of Genome Sciences, University of Washington School of Medicine, Seattle, WA 98195, USA.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Ramage
authors.affiliation: Departments of Medicine and Microbiology, University of Washington School of Medicine, Seattle, WA 98195, USA.
authors.forename: Pradeep K
authors.initials: PK
authors.lastname: Singh
authors.affiliation: Department of Genome Sciences, University of Washington School of Medicine, Seattle, WA 98195, USA.
authors.forename: Colin
authors.initials: C
authors.lastname: Manoil
authors.affiliation: Department of Genome Sciences, University of Washington School of Medicine, Seattle, WA 98195, USA; Departments of Medicine and Microbiology, University of Washington School of Medicine, Seattle, WA 98195, USA; Department of Genome Sciences, University of Washington School of Medicine, 850 Republican Street, Brotman Building, Room 154, Seattle, WA 98109, USA. Electronic address: jimbruce@u.washington.edu.
authors.forename: James E
authors.initials: JE
authors.lastname: Bruce
mesh_terms: D000151:Acinetobacter Infections / Q000382:microbiology* / Q000503:physiopathology*; D040981:Acinetobacter baumannii / Q000502:physiology*; D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000378:metabolism; D054884:Host-Pathogen Interactions; D006801:Humans; D051379:Mice; D018613:Microscopy, Confocal; D054730:Protein Interaction Domains and Motifs; D017434:Protein Structure, Tertiary; D037521:Virulence Factors / Q000737:chemistry / Q000378:metabolism
chemical_list: D001425:Bacterial Outer Membrane Proteins; D037521:Virulence Factors; C078167:OMPA outer membrane proteins