The ability of an attaching and effacing pathogen to trigger localized actin assembly contributes to virulence by promoting mucosal attachment

Enterohaemorrhagic Escherichia coli (EHEC) colonizes the intestine and causes bloody diarrhoea and kidney failure by producing Shiga toxin. Upon binding intestinal cells, EHEC triggers a change in host cell shape, generating actin ‘pedestals’ beneath bound bacteria. To investigate the importance of pedestal formation to disease, we infected genetically engineered mice incapable of supporting pedestal formation by an EHEC‐like mouse pathogen, or wild type mice with a mutant of that pathogen incapable of generating pedestals. We found that pedestal formation promotes attachment of bacteria to the intestinal mucosa and vastly increases the severity of Shiga toxin‐mediated disease.

[1]  V. Vanguri,et al.  A novel murine infection model for Shiga toxin-producing Escherichia coli. , 2012, The Journal of clinical investigation.

[2]  G. Núñez,et al.  Regulated Virulence Controls the Ability of a Pathogen to Compete with the Gut Microbiota , 2012, Science.

[3]  H. De Greve,et al.  O157:H7 and O104:H4 Vero/Shiga toxin-producing Escherichia coli outbreaks: respective role of cattle and humans , 2012, Veterinary Research.

[4]  L. Beutin,et al.  Outbreak of Shiga toxin-producing Escherichia coli (STEC) O104:H4 infection in Germany causes a paradigm shift with regard to human pathogenicity of STEC strains. , 2012, Journal of food protection.

[5]  S. Tzipori,et al.  Allele- and Tir-Independent Functions of Intimin in Diverse Animal Infection Models , 2012, Front. Microbio..

[6]  L. Teel,et al.  Pathogenesis of Shiga-toxin producing escherichia coli. , 2012, Current topics in microbiology and immunology.

[7]  A. Mellmann,et al.  Characterisation of the Escherichia coli strain associated with an outbreak of haemolytic uraemic syndrome in Germany, 2011: a microbiological study. , 2011, The Lancet. Infectious diseases.

[8]  James H. Bullard,et al.  Origins of the E. coli strain causing an outbreak of hemolytic-uremic syndrome in Germany. , 2011, The New England journal of medicine.

[9]  S. Schüller.  Shiga Toxin Interaction with Human Intestinal Epithelium , 2011, Toxins.

[10]  A. O’Brien,et al.  Mouse Models of Escherichia coli O157:H7 Infection and Shiga Toxin Injection , 2011, Journal of biomedicine & biotechnology.

[11]  T. Obrig.  Escherichia coli Shiga Toxin Mechanisms of Action in Renal Disease , 2010, Toxins.

[12]  A. Maldonado-Contreras,et al.  Salmonella Pathogenesis and Processing of Secreted Effectors by Caspase-3 , 2010, Science.

[13]  Paul Dean,et al.  The EspF Effector, a Bacterial Pathogen's Swiss Army Knife , 2010, Infection and Immunity.

[14]  S. Snapper,et al.  Enterohemorrhagic E. coli Requires N-WASP for Efficient Type III Translocation but Not for EspFU-Mediated Actin Pedestal Formation , 2010, PLoS pathogens.

[15]  K. Campellone,et al.  Cytoskeleton‐modulating effectors of enteropathogenic and enterohaemorrhagic Escherichia coli: Tir, EspFU and actin pedestal assembly , 2010, The FEBS journal.

[16]  S. Snapper,et al.  Neural Wiskott-Aldrich syndrome protein modulates Wnt signaling and is required for hair follicle cycling in mice. , 2010, The Journal of clinical investigation.

[17]  V. Gannon,et al.  Verocytotoxin-producing Escherichia coli (VTEC). , 2010, Veterinary microbiology.

[18]  A. Mousnier,et al.  Dissecting the role of the Tir:Nck and Tir:IRTKS/IRSp53 signalling pathways in vivo , 2009, Molecular microbiology.

[19]  B. Finlay,et al.  Molecular mechanisms of Escherichia coli pathogenicity , 2012, Nature Reviews Microbiology.

[20]  John Cowden,et al.  Escherichia coli O157 , 2009, BMJ : British Medical Journal.

[21]  Pamela S. Robinson,et al.  Establishment of conditionally immortalized epithelial cell lines from the intestinal tissue of adult normal and transgenic mice. , 2009, American journal of physiology. Gastrointestinal and liver physiology.

[22]  G. Frankel,et al.  Modelling of Infection by Enteropathogenic Escherichia coli Strains in Lineages 2 and 4 Ex Vivo and In Vivo by Using Citrobacter rodentium Expressing TccP , 2009, Infection and Immunity.

[23]  S. Satchell,et al.  Shiga Toxin 2 Targets the Murine Renal Collecting Duct Epithelium , 2009, Infection and Immunity.

[24]  Hui-Chun Cheng,et al.  Repetitive N-WASP–Binding Elements of the Enterohemorrhagic Escherichia coli Effector EspFU Synergistically Activate Actin Assembly , 2008, PLoS pathogens.

[25]  Responses of Cattle to Gastrointestinal Colonization by Escherichia coli O157:H7 , 2008, Infection and Immunity.

[26]  L. Zimmerhackl,et al.  Treatment and outcome of Shiga-toxin-associated hemolytic uremic syndrome (HUS) , 2008, Pediatric Nephrology.

[27]  M. Troxell,et al.  Mouse Model of Hemolytic-Uremic Syndrome Caused by Endotoxin-Free Shiga Toxin 2 (Stx2) and Protection from Lethal Outcome by Anti-Stx2 Antibody , 2008, Infection and Immunity.

[28]  J. Nougayrède,et al.  EspF Interacts with Nucleation-Promoting Factors To Recruit Junctional Proteins into Pedestals for Pedestal Maturation and Disruption of Paracellular Permeability , 2008, Infection and Immunity.

[29]  S. Tzipori,et al.  EspFU, a type III‐translocated effector of actin assembly, fosters epithelial association and late‐stage intestinal colonization by E. coli O157:H7 , 2008, Cellular microbiology.

[30]  G. Frankel,et al.  Attaching effacing Escherichia coli and paradigms of Tir‐triggered actin polymerization: getting off the pedestal , 2008, Cellular microbiology.

[31]  D. Borenshtein,et al.  Utility of the Citrobacter rodentium infection model in laboratory mice , 2008, Current opinion in gastroenterology.

[32]  R. Xavier,et al.  Wiskott–Aldrich syndrome protein (WASP) and N-WASP are critical for T cell development , 2007, Proceedings of the National Academy of Sciences.

[33]  Megha Ghildiyal,et al.  Enterohaemorrhagic and enteropathogenic Escherichia coli Tir proteins trigger a common Nck‐independent actin assembly pathway , 2007, Cellular microbiology.

[34]  B. Kenny,et al.  Tir phosphorylation and Nck/N‐WASP recruitment by enteropathogenic and enterohaemorrhagic Escherichia coli during ex vivo colonization of human intestinal mucosa is different to cell culture models , 2007, Cellular microbiology.

[35]  J. Kaper,et al.  Shiga toxin binding in normal and inflamed human intestinal mucosa. , 2007, Microbes and infection.

[36]  T. Obrig,et al.  A murine model of HUS: Shiga toxin with lipopolysaccharide mimics the renal damage and physiologic response of human disease. , 2006, Journal of the American Society of Nephrology : JASN.

[37]  A. Abe,et al.  Enteropathogenic Escherichia coli, Shigella flexneri, and Listeria monocytogenes Recruit a Junctional Protein, Zonula Occludens-1, to Actin Tails and Pedestals , 2006, Infection and Immunity.

[38]  Tetsuya Hayashi,et al.  TccP2 of O157:H7 and Non-O157 Enterohemorrhagic Escherichia coli (EHEC): Challenging the Dogma of EHEC-Induced Actin Polymerization , 2006, Infection and Immunity.

[39]  B. Finlay,et al.  Evidence that Tight Junctions Are Disrupted Due to Intimate Bacterial Contact and Not Inflammation during Attaching and Effacing Pathogen Infection In Vivo , 2006, Infection and Immunity.

[40]  M. J. Smith,et al.  Shiga toxin of enterohemorrhagic Escherichia coli type O157:H7 promotes intestinal colonization. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[41]  A. Whale,et al.  A novel category of enteropathogenic Escherichia coli simultaneously utilizes the Nck and TccP pathways to induce actin remodelling , 2006, Cellular microbiology.

[42]  J. Leong,et al.  Exploiting pathogenic Escherichia coli to model transmembrane receptor signalling , 2006, Nature Reviews Microbiology.

[43]  E. Caron,et al.  Subversion of actin dynamics by EPEC and EHEC. , 2006, Current opinion in microbiology.

[44]  M. Woodward,et al.  Role of Intimin-Tir Interactions and the Tir-Cytoskeleton Coupling Protein in the Colonization of Calves and Lambs by Escherichia coli O157:H7 , 2006, Infection and Immunity.

[45]  M. Woodward,et al.  Characterization of Two Non-Locus of Enterocyte Effacement-Encoded Type III-Translocated Effectors, NleC and NleD, in Attaching and Effacing Pathogens , 2005, Infection and Immunity.

[46]  G. Frankel,et al.  Enteropathogenic and Enterohemorrhagic Escherichia coli Infections: Translocation, Translocation, Translocation , 2005, Infection and Immunity.

[47]  J. Leong,et al.  Nck‐independent actin assembly is mediated by two phosphorylated tyrosines within enteropathogenic Escherichia coli Tir , 2005, Molecular microbiology.

[48]  P. Tarr,et al.  Shiga-toxin-producing Escherichia coli and haemolytic uraemic syndrome , 2005, The Lancet.

[49]  M. Waldor,et al.  The Locus of Enterocyte Effacement-Encoded Effector Proteins All Promote Enterohemorrhagic Escherichia coli Pathogenicity in Infant Rabbits , 2005, Infection and Immunity.

[50]  Nathan Christopher Shaner,et al.  Actin and alpha-actinin dynamics in the adhesion and motility of EPEC and EHEC on host cells. , 2005, Cell motility and the cytoskeleton.

[51]  É. Oswald,et al.  TccP is an enterohaemorrhagic Escherichia coli O157:H7 type III effector protein that couples Tir to the actin‐cytoskeleton † , 2004, Cellular microbiology.

[52]  S. Clare,et al.  Organ specificity, colonization and clearance dynamics in vivo following oral challenges with the murine pathogen Citrobacter rodentium , 2004, Cellular microbiology.

[53]  J. Leong,et al.  EspFU is a translocated EHEC effector that interacts with Tir and N-WASP and promotes Nck-independent actin assembly. , 2004, Developmental cell.

[54]  D. Kalman,et al.  Enteropathogenic Escherichia coli use redundant tyrosine kinases to form actin pedestals. , 2004, Molecular biology of the cell.

[55]  Daniel Metzger,et al.  Tissue‐specific and inducible Cre‐mediated recombination in the gut epithelium , 2004, Genesis.

[56]  R. Hayward,et al.  Phosphorylation of the enteropathogenic E. coli receptor by the Src-family kinase c-Fyn triggers actin pedestal formation , 2004, Nature Cell Biology.

[57]  S. Clare,et al.  Identification of a Novel Citrobacter rodentium Type III Secreted Protein, EspI, and Roles of This and Other Secreted Proteins in Infection , 2004, Infection and Immunity.

[58]  T. Pawson,et al.  Dissecting virulence: systematic and functional analyses of a pathogenicity island. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[59]  T. Pawson,et al.  Clustering of Nck by a 12-residue Tir phosphopeptide is sufficient to trigger localized actin assembly , 2004, The Journal of cell biology.

[60]  Harry L. T. Mobley,et al.  Pathogenic Escherichia coli , 2004, Nature Reviews Microbiology.

[61]  B. Mayer,et al.  Inducible Clustering of Membrane-Targeted SH3 Domains of the Adaptor Protein Nck Triggers Localized Actin Polymerization , 2004, Current Biology.

[62]  J. Chiu,et al.  Site-directed, Ligase-Independent Mutagenesis (SLIM): a single-tube methodology approaching 100% efficiency in 4 h. , 2004, Nucleic acids research.

[63]  M. Waldor,et al.  Critical Roles for stx2,eae, and tir in EnterohemorrhagicEscherichia coli-Induced Diarrhea and Intestinal Inflammation in Infant Rabbits , 2003, Infection and Immunity.

[64]  S. Clare,et al.  Central Role for B Lymphocytes and CD4+ T Cells in Immunity to Infection by the Attaching and Effacing Pathogen Citrobacter rodentium , 2003, Infection and Immunity.

[65]  P. Desmarchelier,et al.  Presence of Activatable Shiga Toxin Genotype (stx2d) in Shiga Toxigenic Escherichia coli from Livestock Sources , 2003, Journal of Clinical Microbiology.

[66]  J. Nougayrède,et al.  Adhesion of enteropathogenic Escherichia coli to host cells , 2003, Cellular microbiology.

[67]  B. Finlay,et al.  Citrobacter rodentium translocated intimin receptor (Tir) is an essential virulence factor needed for actin condensation, intestinal colonization and colonic hyperplasia in mice , 2003, Molecular microbiology.

[68]  S. Yokoyama,et al.  Adhesion and Colonization of Enterohemorrhagic Escherichia coli O157:H7 in Cecum of Mice , 2003, Microbiology and immunology.

[69]  B. Finlay,et al.  Mice Lacking T and B Lymphocytes Develop Transient Colitis and Crypt Hyperplasia yet Suffer Impaired Bacterial Clearance during Citrobacter rodentium Infection , 2002, Infection and Immunity.

[70]  J. Leong,et al.  A tyrosine‐phosphorylated 12‐amino‐acid sequence of enteropathogenic Escherichia coli Tir binds the host adaptor protein Nck and is required for Nck localization to actin pedestals , 2002, Molecular microbiology.

[71]  M. Goldberg.  Actin-Based Motility of Intracellular Microbial Pathogens , 2001, Microbiology and Molecular Biology Reviews.

[72]  H Schmidt,et al.  Shiga-toxin-converting bacteriophages. , 2001, Research in microbiology.

[73]  Sheila M. Thomas,et al.  N-WASP deficiency reveals distinct pathways for cell surface projections and microbial actin-based motility , 2001, Nature Cell Biology.

[74]  M. Woodward,et al.  Attaching and effacing lesions caused by Escherichia coli O157:H7 in experimentally inoculated neonatal lambs. , 2001, Journal of medical microbiology.

[75]  T. Pawson,et al.  Enteropathogenic E. coli Tir binds Nck to initiate actin pedestal formation in host cells , 2001, Nature Cell Biology.

[76]  G. Dougan,et al.  Intimin-Specific Immune Responses Prevent Bacterial Colonization by the Attaching-Effacing PathogenCitrobacter rodentium , 2001, Infection and Immunity.

[77]  K. Rottner,et al.  Actin pedestal formation by enteropathogenic Escherichia coli and intracellular motility of Shigella flexneri are abolished in N‐WASP‐defective cells , 2001, EMBO reports.

[78]  M. Kirschner,et al.  Nck and Phosphatidylinositol 4,5-Bisphosphate Synergistically Activate Actin Polymerization through the N-WASP-Arp2/3 Pathway* , 2001, The Journal of Biological Chemistry.

[79]  J. Nougayrède,et al.  Translocated EspF protein from enteropathogenic Escherichia coli disrupts host intestinal barrier function. , 2001, The Journal of clinical investigation.

[80]  T. Takeda,et al.  Escherichia coli Shiga toxin. , 2000, Journal of natural toxins.

[81]  J. Nougayrède,et al.  Role of Tir and Intimin in the Virulence of Rabbit Enteropathogenic Escherichia coli Serotype O103:H2 , 2000, Infection and Immunity.

[82]  B. Finlay,et al.  Identification of the intimin‐binding domain of Tir of enteropathogenic Escherichia coli , 1999, Cellular microbiology.

[83]  I. Connerton,et al.  Binding of intimin from enteropathogenic Escherichia coli to Tir and to host cells , 1999, Molecular microbiology.

[84]  B. Kenny.  Phosphorylation of tyrosine 474 of the enteropathogenic Escherichia coli (EPEC) Tir receptor molecule is essential for actin nucleating activity and is preceded by additional host modifications , 1999, Molecular microbiology.

[85]  B. Bosworth,et al.  Pathogenesis of Escherichia coli O157:H7 in weaned calves. , 1999, Advances in experimental medicine and biology.

[86]  B. Finlay,et al.  Enteropathogenic E. coli (EPEC) Transfers Its Receptor for Intimate Adherence into Mammalian Cells , 1997, Cell.

[87]  J. Kaper,et al.  Novel form of actin-based motility transports bacteria on the surfaces of infected cells. , 1996, Cell motility and the cytoskeleton.

[88]  F. Gunzer,et al.  The role of the eaeA gene in diarrhea and neurological complications in a gnotobiotic piglet model of enterohemorrhagic Escherichia coli infection , 1995, Infection and immunity.

[89]  T. McDaniel,et al.  A genetic locus of enterocyte effacement conserved among diverse enterobacterial pathogens. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[90]  S. Falkow,et al.  The eae gene of Citrobacter freundii biotype 4280 is necessary for colonization in transmissible murine colonic hyperplasia , 1993, Infection and immunity.

[91]  M. Levine,et al.  Role of the eaeA gene in experimental enteropathogenic Escherichia coli infection. , 1993, The Journal of clinical investigation.

[92]  S. Falkow,et al.  Attaching and effacing locus of a Citrobacter freundii biotype that causes transmissible murine colonic hyperplasia , 1993, Infection and immunity.

[93]  E. Boedeker,et al.  Regional Differences in Attachment of Enteroadherent Escherichia coli Strain RDEC‐1 to Rabbit Intestine: Luminal Colonization But Lack of Mucosal Adherence in Jejunal Self‐Filling Blind Loops , 1987, Journal of pediatric gastroenterology and nutrition.

[94]  M. Doyle,et al.  Colonization of chicken cecae by Escherichia coli associated with hemorrhagic colitis , 1985, Applied and environmental microbiology.

[95]  M. Levine,et al.  Attaching and effacing activities of rabbit and human enteropathogenic Escherichia coli in pig and rabbit intestines , 1983, Infection and immunity.