Enteropathogenic Escherichia coli Activates Ezrin, Which Participates in Disruption of Tight Junction Barrier Function

ABSTRACT Enteropathogenic Escherichia coli (EPEC) is an important human intestinal pathogen, especially in infants. EPEC adherence to intestinal epithelial cells induces the accumulation of a number of cytoskeletal proteins beneath the bacteria, including the membrane-cytoskeleton linker ezrin. Evidence suggests that ezrin can participate in signal transduction. The aim of this study was to determine whether ezrin is activated following EPEC infection and if it is involved in the cross talk with host intestinal epithelial cells. We show here that following EPEC attachment to intestinal epithelial cells there was significant phosphorylation of ezrin, first on threonine and later on tyrosine residues. A significant increase in cytoskeleton-associated ezrin occurred following phosphorylation, suggesting activation of this molecule. Nonpathogenic E. coli and EPEC strains harboring mutations in type III secretion failed to elicit this response. Expression of dominant-negative ezrin significantly decreased the EPEC-elicited association of ezrin with the cytoskeleton and attenuated the disruption of intestinal epithelial tight junctions. These results suggest that ezrin is involved in transducing EPEC-initiated signals that ultimately affect host physiological functions.

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

[2]  E. Benz,et al.  Characterization of the Interaction between Protein 4.1R and ZO-2 , 2000, The Journal of Biological Chemistry.

[3]  J. Rosenberg,et al.  Enteropathogenic Escherichia coli dephosphorylates and dissociates occludin from intestinal epithelial tight junctions , 2000, Cellular microbiology.

[4]  A. Gautreau,et al.  Morphogenic Effects of Ezrin Require a Phosphorylation-Induced Transition from Oligomers to Monomers at the Plasma Membrane , 2000, The Journal of cell biology.

[5]  P. Karplus,et al.  Structure of the ERM Protein Moesin Reveals the FERM Domain Fold Masked by an Extended Actin Binding Tail Domain , 2000, Cell.

[6]  E. Rodriguez-Boulan,et al.  Ezrin Promotes Morphogenesis of Apical Microvilli and Basal Infoldings in Retinal Pigment Epithelium , 1999, The Journal of cell biology.

[7]  J Mounier,et al.  A functional role for ezrin during Shigella flexneri entry into epithelial cells. , 1999, Journal of cell science.

[8]  T. Matsui,et al.  Immunofluorescence detection of ezrin/radixin/moesin (ERM) proteins with their carboxyl-terminal threonine phosphorylated in cultured cells and tissues. , 1999, Journal of cell science.

[9]  M. Schmidt,et al.  Insertion of EspD into epithelial target cell membranes by infecting enteropathogenic Escherichia coli , 1999, Molecular microbiology.

[10]  G. Hecht,et al.  Enteropathogenic E. coli attenuates secretagogue-induced net intestinal ion transport but not Cl- secretion. , 1999, American journal of physiology. Gastrointestinal and liver physiology.

[11]  A. Bretscher,et al.  C-terminal threonine phosphorylation activates ERM proteins to link the cell's cortical lipid bilayer to the cytoskeleton. , 1998, Biochemical and biophysical research communications.

[12]  M. Arpin,et al.  Effect of hepatocyte growth factor on assembly of zonula occludens‐1 protein at the plasma membrane , 1998, Journal of cellular physiology.

[13]  M. Rohde,et al.  Small GTP-binding proteins of the Rho- and Ras-subfamilies are not involved in the actin rearrangements induced by attaching and effacing Escherichia coli. , 1998, FEMS microbiology letters.

[14]  K. Kaibuchi,et al.  Regulation of Cytoskeleton and Cell Adhesions by the Small GTPase Rho and Its Targets. , 1998, Trends in cardiovascular medicine.

[15]  K. Hahn,et al.  Agents That Inhibit Rho, Rac, and Cdc42 Do Not Block Formation of Actin Pedestals in HeLa Cells Infected with Enteropathogenic Escherichia coli , 1998, Infection and Immunity.

[16]  T. Baldwin.  Pathogenicity of enteropathogenic Escherichia coli , 1998 .

[17]  N. Takahashi,et al.  Ezrin/Radixin/Moesin (ERM) Proteins Bind to a Positively Charged Amino Acid Cluster in the Juxta-Membrane Cytoplasmic Domain of CD44, CD43, and ICAM-2 , 1998, The Journal of cell biology.

[18]  K. Kaibuchi,et al.  Rho-Kinase Phosphorylates COOH-terminal Threonines of Ezrin/Radixin/Moesin (ERM) Proteins and Regulates Their Head-to-Tail Association , 1998, The Journal of cell biology.

[19]  S. Knutton,et al.  Rapid modulation of electrolyte transport in Caco-2 cell monolayers by enteropathogenic Escherichia coli (EPEC) infection , 1998, Gut.

[20]  A. Bretscher,et al.  Ezrin: a protein requiring conformational activation to link microfilaments to the plasma membrane in the assembly of cell surface structures. , 1997, Journal of cell science.

[21]  S. Savkovic,et al.  Enteropathogenic Escherichia coli-induced myosin light chain phosphorylation alters intestinal epithelial permeability. , 1997, Gastroenterology.

[22]  Shoichiro Tsukita,et al.  ERM (Ezrin/Radixin/Moesin)-based Molecular Mechanism of Microvillar Breakdown at an Early Stage of Apoptosis , 1997, The Journal of cell biology.

[23]  A. Bretscher,et al.  Identification of EBP50: A PDZ-containing Phosphoprotein that Associates with Members of the Ezrin-Radixin-Moesin Family , 1997, The Journal of cell biology.

[24]  J. Jääskeläinen,et al.  The ezrin protein family: membrane-cytoskeleton interactions and disease associations. , 1997, Current opinion in cell biology.

[25]  S. Savkovic,et al.  Activation of NF-κB in intestinal epithelial cells by enteropathogenic Escherichia coli. , 1997, American journal of physiology. Cell physiology.

[26]  A. Hall,et al.  Rho- and Rac-dependent Assembly of Focal Adhesion Complexes and Actin Filaments in Permeabilized Fibroblasts: An Essential Role for Ezrin/Radixin/Moesin Proteins , 1997, The Journal of cell biology.

[27]  J. Oh,et al.  Activation of host cell protein kinase C by enteropathogenic Escherichia coli , 1997, Infection and immunity.

[28]  Alexis Gautreau,et al.  Ezrin Is an Effector of Hepatocyte Growth Factor–mediated Migration and Morphogenesis in Epithelial Cells , 1997, The Journal of cell biology.

[29]  S. Tsukita,et al.  ERM proteins: head-to-tail regulation of actin-plasma membrane interaction. , 1997, Trends in biochemical sciences.

[30]  S. Savkovic,et al.  Attachment of a noninvasive enteric pathogen, enteropathogenic Escherichia coli, to cultured human intestinal epithelial monolayers induces transmigration of neutrophils , 1996, Infection and immunity.

[31]  P. Williams,et al.  Phosphorylation of myosin light chain at distinct sites and its association with the cytoskeleton during enteropathogenic Escherichia coli infection , 1996, Infection and immunity.

[32]  P. Williams,et al.  Hijacking host cell signal transduction mechanisms during infection with enteropathogenic Escherichia coli. , 1996, Biochemical Society transactions.

[33]  D. Philpott,et al.  Infection of T84 cells with enteropathogenic Escherichia coli alters barrier and transport functions. , 1996, The American journal of physiology.

[34]  T. Nakamura,et al.  Induction of tyrosine phosphorylation and translocation of ezrin by hepatocyte growth factor/scatter factor. , 1995, Biochemical and biophysical research communications.

[35]  T. McDaniel,et al.  Enteropathogenic Escherichia coli contains a putative type III secretion system necessary for the export of proteins involved in attaching and effacing lesion formation. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[36]  J. Cohn,et al.  Dephosphorylation of ezrin as an early event in renal microvillar breakdown and anoxic injury. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[37]  J. Alverdy,et al.  Enteropathogenic Escherichia coli adherence to intestinal epithelial monolayers diminishes barrier function. , 1995, The American journal of physiology.

[38]  A. Vaheri,et al.  Ezrin has a COOH-terminal actin-binding site that is conserved in the ezrin protein family , 1994, The Journal of cell biology.

[39]  N. Sato,et al.  ERM family members as molecular linkers between the cell surface glycoprotein CD44 and actin-based cytoskeletons , 1994, The Journal of cell biology.

[40]  B. Finlay,et al.  The eaeB gene of enteropathogenic Escherichia coli is necessary for signal transduction in epithelial cells , 1994, Infection and immunity.

[41]  E. Rosen,et al.  Hepatocyte growth factor/scatter factor effects on epithelia. Regulation of intercellular junctions in transformed and nontransformed cell lines, basolateral polarization of c-met receptor in transformed and natural intestinal epithelia, and induction of rapid wound repair in a transformed model epi , 1994, The Journal of clinical investigation.

[42]  P. Sherman,et al.  Signal transduction in human epithelial cells infected with attaching and effacing Escherichia coli in vitro. , 1994, Gastroenterology.

[43]  A. Bretscher,et al.  Ezrin is concentrated in the apical microvilli of a wide variety of epithelial cells whereas moesin is found primarily in endothelial cells. , 1993, Journal of cell science.

[44]  B. Finlay,et al.  Enteropathogenic Escherichia coli decreases the transepithelial electrical resistance of polarized epithelial monolayers , 1993, Infection and immunity.

[45]  M. Arpin,et al.  Ezrin contains cytoskeleton and membrane binding domains accounting for its proposed role as a membrane-cytoskeletal linker , 1993, The Journal of cell biology.

[46]  S. Colgan,et al.  A simple approach to measurement of electrical parameters of cultured epithelial monolayers: Use in assessing neutrophil-epithelial interactions , 1992 .

[47]  B. Finlay,et al.  Signal transduction between enteropathogenic Escherichia coli (EPEC) and epithelial cells: EPEC induces tyrosine phosphorylation of host cell proteins to initiate cytoskeletal rearrangement and bacterial uptake. , 1992, The EMBO journal.

[48]  T. Hunter,et al.  Identification of the two major epidermal growth factor-induced tyrosine phosphorylation sites in the microvillar core protein ezrin. , 1992, The Journal of biological chemistry.

[49]  N. Sato,et al.  A gene family consisting of ezrin, radixin and moesin. Its specific localization at actin filament/plasma membrane association sites. , 1992, Journal of cell science.

[50]  B. Finlay,et al.  Cytoskeletal composition of attaching and effacing lesions associated with enteropathogenic Escherichia coli adherence to HeLa cells , 1992, Infection and immunity.

[51]  P. Williams,et al.  Purification of a 20 kDa phosphoprotein from epithelial cells and identification as a myosin light chain Phosphorylation induced by enteropathogenicEscherichia coli and phorbol ester , 1991, FEBS letters.

[52]  A. Bretscher,et al.  The secretion‐stimulated 80K phosphoprotein of parietal cells is ezrin, and has properties of a membrane cytoskeletal linker in the induced apical microvilli. , 1991, The EMBO journal.

[53]  W. Pfaller,et al.  Morphology of the differentiation and maturation of LLC‐PK1 epithelia , 1990, Journal of cellular physiology.

[54]  A. Bretscher.  Rapid phosphorylation and reorganization of ezrin and spectrin accompany morphological changes induced in A-431 cells by epidermal growth factor , 1989, The Journal of cell biology.

[55]  J. Madara,et al.  Structural analysis of a human intestinal epithelial cell line. , 1987, Gastroenterology.

[56]  S. Knutton,et al.  Adhesion of enteropathogenic Escherichia coli to human intestinal enterocytes and cultured human intestinal mucosa , 1987, Infection and immunity.

[57]  T. Kreis.  Microinjected antibodies against the cytoplasmic domain of vesicular stomatitis virus glycoprotein block its transport to the cell surface. , 1986, The EMBO journal.

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

[59]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[60]  S. Tsukita,et al.  ERM (ezrin/radixin/moesin) family: from cytoskeleton to signal transduction. , 1997, Current opinion in cell biology.

[61]  E. Rosen,et al.  Hepatocyte Growth Factor / Scatter Factor Effects on Epithelia , 2022 .