title: EspT triggers formation of lamellipodia and membrane ruffles through activation of Rac-1 and Cdc42.
abstract: Subversion of the eukaryotic cell cytoskeleton is a virulence strategy employed by many bacterial pathogens. Due to the pivotal role of Rho GTPases in actin dynamics they are common targets of bacterial effector proteins and toxins. IpgB1, IpgB2 (Shigella), SifA, SifB (Salmonella) and Map and EspM (attaching and effacing pathogens) constitute a family of type III secretion system effectors that subverts small GTPase signalling pathways. In this study we identified and characterized EspT from Citrobacter rodentium that triggers formation of lamellipodia on Swiss 3T3 and membrane ruffles on HeLa cells, which are reminiscent of the membrane ruffles induced by IpgB1. Ectopic expression of EspT and IpgB1, but not EspM, resulted in a mitochondrial localization. Using dominant negative constructs we found that EspT-induced actin remodelling is dependent on GTP-bound Rac-1 and Cdc42 but not ELMO or Dock180, which are hijacked by IpgB1 in order to form a Rac-1 specific guanine nucleotide exchange factor. Using pull-down assays with the Rac-1 and Cdc42 binding domains of Pak and WASP we demonstrate that EspT is capable of activating both Rac-1 and Cdc42. These results suggest that EspT modulates the host cell cytoskeleton through coactivation of Rac-1 and Cdc42 by a distinct mechanism.
authors.affiliation: Centre for Molecular Microbiology and Infection, Division of Cell and Molecular Biology, Imperial College, London SW7 2AZ, UK.
authors.forename: Richard R
authors.initials: RR
authors.lastname: Bulgin
authors.forename: Ana
authors.initials: A
authors.lastname: Arbeloa
authors.forename: Jade C S
authors.initials: JC
authors.lastname: Chung
authors.forename: Gad
authors.initials: G
authors.lastname: Frankel
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000382:microbiology*; D044082:Citrobacter rodentium / Q000502:physiology*; D004847:Epithelial Cells / Q000382:microbiology; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D011485:Protein Binding; D011554:Pseudopodia / Q000382:microbiology*; D016415:Sequence Alignment; D037521:Virulence Factors / Q000378:metabolism*; D020764:cdc42 GTP-Binding Protein / Q000378:metabolism*; D020830:rac1 GTP-Binding Protein / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D037521:Virulence Factors; D020764:cdc42 GTP-Binding Protein; D020830:rac1 GTP-Binding Protein