title: Enteropathogenic Escherichia coli dynamically regulates EGFR signaling in intestinal epithelial cells.
abstract: The diarrheagenic pathogen enteropathogenic Escherichia coli (EPEC) dynamically modulates the survival of infected host intestinal epithelial cells. In the initial stages of infection, several prosurvival signaling events are activated in host cells. These include the phosphorylation of epidermal growth factor receptor (EGFR) and the consequent activation of the phosphatidylinositol-3 kinase/Akt pathway. While studying this pathway in infected epithelial cells, we observed EGFR depletion at later stages of infection, followed subsequently by a decrease in phospho-EGFR. EGFR loss was not dependent on receptor phosphorylation, or on canonical proteasome- and lysosome-dependent processes. Although a type III secretion mutant (ΔescN) stimulated EGFR phosphorylation, it failed to induce receptor degradation. To identify the specific EPEC effector molecule(s) that influenced EGFR stability, epithelial cells infected with isogenic mutant EPEC strains were examined. An EPEC ΔespF strain failed to induce EGFR degradation, whereas EPEC ΔespZ accentuated receptor loss in infected cells. Given the known and contrasting effects of EspF and EspZ on caspase activation, and the known role of proteases in cleaving EGFR, we explored the effect of caspase inhibitors on infection-dependent EGFR loss. The pan-caspase inhibitor Q-VD-OPh blocked EPEC-induced EGFR cleavage in a dose-dependent manner. Taken together, our data suggest that EPEC EspF stimulates caspase-dependent EGFR cleavage and loss, whereas EspZ inhibits this process. Whereas EGFR phosphorylation contributes to the survival of host cells early in infection, EspF-driven caspase activation and consequent EGFR loss likely induce a precipitous increase in host cell death later in the infectious process.
authors.affiliation: School of Animal and Comparative Biomedical Sciences, University of Arizona, Tucson, Arizona;
authors.forename: Jennifer Lising
authors.initials: JL
authors.lastname: Roxas
authors.affiliation: School of Animal and Comparative Biomedical Sciences, University of Arizona, Tucson, Arizona;
authors.forename: Katheryn
authors.initials: K
authors.lastname: Ryan
authors.affiliation: School of Animal and Comparative Biomedical Sciences, University of Arizona, Tucson, Arizona; Department of Immunobiology, University of Arizona, Tucson, Arizona; and The BIO5 Institute for Collaborative Research, University of Arizona, Tucson, Arizona.
authors.forename: Gayatri
authors.initials: G
authors.lastname: Vedantam
authors.affiliation: School of Animal and Comparative Biomedical Sciences, University of Arizona, Tucson, Arizona; Department of Immunobiology, University of Arizona, Tucson, Arizona; and The BIO5 Institute for Collaborative Research, University of Arizona, Tucson, Arizona vkv@email.arizona.edu.
authors.forename: V K
authors.initials: VK
authors.lastname: Viswanathan
mesh_terms: D018938:Caco-2 Cells; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D061945:Caspase Inhibitors / Q000494:pharmacology; D016923:Cell Death; D004305:Dose-Response Relationship, Drug; D054308:Enteropathogenic Escherichia coli / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism* / Q000382:microbiology* / Q000473:pathology; D066246:ErbB Receptors / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D054884:Host-Pathogen Interactions; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism*; D007422:Intestines / Q000187:drug effects / Q000382:microbiology* / Q000473:pathology; D047908:Intracellular Signaling Peptides and Proteins; D009154:Mutation; D010766:Phosphorylation; D055550:Protein Stability; D015398:Signal Transduction / Q000187:drug effects; D013997:Time Factors
chemical_list: D002352:Carrier Proteins; D061945:Caspase Inhibitors; D029968:Escherichia coli Proteins; C489109:EspFU protein, E coli; C569164:EspZ protein, E coli; D047908:Intracellular Signaling Peptides and Proteins; C512478:EGFR protein, human; D066246:ErbB Receptors
keywords: EGFR degradation; EGFR phosphorylation; EPEC; EspF; EspZ; caspase