title: Autoinducer 3 and epinephrine signaling in the kinetics of locus of enterocyte effacement gene expression in enterohemorrhagic Escherichia coli.
abstract: Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is responsible for causing outbreaks of bloody diarrhea and hemolytic-uremic syndrome throughout the world. The locus of enterocyte effacement (LEE) consists of five major operons and is required for the formation of attaching and effacing lesions that disrupt intestinal epithelial microvilli. We have previously reported that expression of EHEC LEE genes is regulated by the luxS quorum-sensing system. The luxS gene in EHEC affects the production of autoinducer 3 (AI-3), which activates the LEE. Epinephrine and norepinephrine also activate the LEE in a manner similar to that of AI-3. Previous studies of quorum-sensing regulation of LEE transcription have thus far been restricted to using reporter systems in an E. coli K-12 background. Here, we examined the kinetics of LEE gene transcription, protein expression, and function of the LEE type III secretion apparatus in wild-type (WT) EHEC and an isogenic luxS mutant. The results revealed that the luxS mutant had diminished transcription from the LEE promoters during the mid-exponential growth phase; decreased protein levels of EscJ, Tir, and EspA; and reduced secretion of EspA and EspB. The luxS mutation also caused a delay in the formation of attaching and effacing lesions on cultured epithelial cells compared to the wild type. Epinephrine enhanced LEE expression in both the WT and the luxS mutant, but the WT still exhibited greater LEE activation. The results suggest a possible synergistic relationship between AI-3 and epinephrine. The combined effects of these two signaling molecules may lead to greater LEE expression and a more efficient infection.
authors.affiliation: University of Texas Southwestern Medical Center, Dept. of Microbiology, 5323 Harry Hines Blvd., Dallas, TX 75390-9048. Vanessa.Sperandio@UTSouthwestern.edu.
authors.forename: Matthew
authors.initials: M
authors.lastname: Walters
authors.forename: Vanessa
authors.initials: V
authors.lastname: Sperandio
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000502:physiology*; D013437:Carbon-Sulfur Lyases / Q000235:genetics / Q000502:physiology*; D002478:Cells, Cultured; D004837:Epinephrine / Q000494:pharmacology*; D004927:Escherichia coli Infections / Q000382:microbiology; D019453:Escherichia coli O157 / Q000187:drug effects* / Q000235:genetics; D029968:Escherichia coli Proteins / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D007700:Kinetics; D009154:Mutation; D010750:Phosphoproteins / Q000235:genetics*; D015398:Signal Transduction; D014158:Transcription, Genetic / Q000187:drug effects
chemical_list: D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; C457340:LEE protein, E coli; D010750:Phosphoproteins; D013437:Carbon-Sulfur Lyases; C117753:LuxS protein, Bacteria; D004837:Epinephrine