title: Enteropathogenic E. coli effectors EspG1/G2 disrupt microtubules, contribute to tight junction perturbation and inhibit restoration.
abstract: Enteropathogenic Escherichia coli (EPEC) uses a type 3 secretion system to transfer effector proteins into the host intestinal epithelial cell. Several effector molecules contribute to tight junction disruption including EspG1 and its homologue EspG2 via a mechanism thought to involve microtubule destruction. The aim of this study was to investigate the contribution of EspG-mediated microtubule disruption to TJ perturbation. We demonstrate that wild type EPEC infection disassembles microtubules and induces the progressive movement of occludin away from the membrane and into the cytosol. Deletion of espG1/G2 attenuates both of these phenotypes. In addition, EPEC infection impedes barrier recovery from calcium switch, suggesting that inhibition of TJ restoration, not merely disruption, prolongs barrier loss. TJs recover more rapidly following infection with ΔespG1/G2 than with wild type EPEC, demonstrating that EspG1/G2 perpetuate barrier loss. Although EspG regulates ADP-ribosylation factor (ARF) and p21-activated kinase (PAK), these activities are not necessary for microtubule destruction or perturbation of TJ structure and function. These data strongly support a role for EspG1/G2 and its associated effects on microtubules in delaying the recovery of damaged tight junctions caused by EPEC infection.
authors.affiliation: Department of Microbiology & Immunology, University of Illinois at Chicago, 835 S. Wolcott, (M/C 790), Chicago, IL, 60612, USA.
authors.forename: Lila G
authors.initials: LG
authors.lastname: Glotfelty
authors.forename: Anita
authors.initials: A
authors.lastname: Zahs
authors.forename: Kimberley
authors.initials: K
authors.lastname: Hodges
authors.forename: Kuangda
authors.initials: K
authors.lastname: Shan
authors.forename: Neal M
authors.initials: NM
authors.lastname: Alto
authors.forename: Gail A
authors.initials: GA
authors.lastname: Hecht
mesh_terms: D054308:Enteropathogenic Escherichia coli / Q000254:growth & development* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000378:metabolism*; D054884:Host-Pathogen Interactions; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism*; D019108:Tight Junctions / Q000502:physiology*; D037521:Virulence Factors / Q000378:metabolism*
chemical_list: D029968:Escherichia coli Proteins; C000599182:EspG1 protein, E coli; C505458:EspG2 protein, E coli; D008869:Microtubule-Associated Proteins; D037521:Virulence Factors