title: The fibronectin-binding motif within FlpA facilitates Campylobacter jejuni adherence to host cell and activation of host cell signaling.
abstract: Campylobacter jejuni is a gram-negative, curved and rod-shaped bacterium that causes human gastroenteritis. Acute disease is associated with C. jejuni invasion of the intestinal epithelium. Epithelial cells infected with C. jejuni strains containing mutations in the FlpA and CadF fibronectin (Fn)-binding proteins exhibit reduced invasion of host cells and a C. jejuni CadF FlpA double mutant is impaired in the activation of epidermal growth factor receptor (EGFR) and Rho GTPase Rac1. Although these observations establish a role for Fn-binding proteins during C. jejuni invasion, their mechanistic contributions to invasion-associated signaling are unclear. We examined FlpA, a C. jejuni Fn-binding protein composed of three FNIII-like repeats D1, D2 and D3, to identify the interactions required for cellular adherence on pathogen-induced host cell signaling. We report that FlpA binds the Fn gelatin-binding domain via a motif within the D2 repeat. Epithelial cells infected with a flpA mutant exhibited decreased Rac1 activation and reduced membrane ruffling that coincided with impaired delivery of the secreted Cia proteins and reduced cell association. Phosphorylation of the Erk1/2 kinase, a downstream effector of EGFR signaling, was specifically associated with FlpA-mediated activation of β1-integrin and EGFR signaling. In vivo experiments revealed that FlpA is necessary for C. jejuni disease based on bacterial dissemination to the spleen of IL-10(-/-) germ-free mice. Thus, a novel Fn-binding motif within FlpA potentiates activation of Erk1/2 signaling via β1-integrin during C. jejuni infection.
authors.affiliation: School of Molecular Biosciences, College of Veterinary Medicine, Washington State University , Pullman, WA 99164-7523, USA.
authors.forename: Charles L
authors.initials: CL
authors.lastname: Larson
authors.affiliation: School of Molecular Biosciences, College of Veterinary Medicine, Washington State University , Pullman, WA 99164-7523, USA.
authors.forename: Derrick R
authors.initials: DR
authors.lastname: Samuelson
authors.affiliation: School of Molecular Biosciences, College of Veterinary Medicine, Washington State University , Pullman, WA 99164-7523, USA.
authors.forename: Tyson P
authors.initials: TP
authors.lastname: Eucker
authors.affiliation: School of Molecular Biosciences, College of Veterinary Medicine, Washington State University , Pullman, WA 99164-7523, USA.
authors.forename: Jason L
authors.initials: JL
authors.lastname: O'Loughlin
authors.affiliation: School of Molecular Biosciences, College of Veterinary Medicine, Washington State University , Pullman, WA 99164-7523, USA.
authors.forename: Michael E
authors.initials: ME
authors.lastname: Konkel
keywords: MSCRAMM; adherence; adhesion; extracellular matrix; lipoprotein; membrane