title: RcsB determines the locus of enterocyte effacement (LEE) expression and adherence phenotype of Escherichia coli O157 : H7 spinach outbreak strain TW14359 and coordinates bicarbonate-dependent LEE activation with repression of motility.
abstract: The 2006 US spinach outbreak of Escherichia coli O157 : H7, characterized by unusually severe disease, has been attributed to a strain (TW14359) with enhanced pathogenic potential, including elevated virulence gene expression, robust adherence and the presence of novel virulence factors. This study proposes a mechanism for the unique virulence expression and adherence phenotype of this strain, and further expands the role for regulator RcsB in control of the E. coli locus of enterocyte effacement (LEE) pathogenicity island. Proteomic analysis of TW14359 revealed a virulence proteome consistent with previous transcriptome studies that included elevated levels of the LEE regulatory protein Ler and type III secretion system (T3SS) proteins, secreted T3SS effectors and Shiga toxin 2. Basal levels of the LEE activator and Rcs phosphorelay response regulator, RcsB, were increased in strain TW14359 relative to O157 : H7 strain Sakai. Deletion of rcsB eliminated inherent differences between these strains in ler expression, and in T3SS-dependent adherence. A reciprocating regulatory pathway involving RcsB and LEE-encoded activator GrlA was identified and predicted to co-ordinate LEE activation with repression of the flhDC flagellar regulator and motility. Overexpression of grlA was shown to increase RcsB levels, but did not alter expression from promoters driving rcsB transcription. Expression of rcsDB and RcsB was determined to increase in response to physiological levels of bicarbonate, and bicarbonate-dependent stimulation of the LEE was shown to be dependent on an intact Rcs system and ler activator grvA. The results of this study significantly broaden the role for RcsB in enterohaemorrhagic E. coli virulence regulation.
authors.affiliation: Department of Cell Biology, Microbiology, and Molecular Biology, University of South Florida, Tampa, FL 33620, USA.
authors.forename: Jason K
authors.initials: JK
authors.lastname: Morgan
authors.affiliation: Department of Cell Biology, Microbiology, and Molecular Biology, University of South Florida, Tampa, FL 33620, USA.
authors.forename: Khoury W
authors.initials: KW
authors.lastname: Vendura
authors.affiliation: Department of Cell Biology, Microbiology, and Molecular Biology, University of South Florida, Tampa, FL 33620, USA.
authors.forename: Stanley M
authors.initials: SM
authors.lastname: Stevens
authors.affiliation: Department of Cell Biology, Microbiology, and Molecular Biology, University of South Florida, Tampa, FL 33620, USA.
authors.forename: James T
authors.initials: JT
authors.lastname: Riordan
mesh_terms: D001422:Bacterial Adhesion; D001639:Bicarbonates / Q000378:metabolism*; D004196:Disease Outbreaks; D004927:Escherichia coli Infections / Q000453:epidemiology / Q000382:microbiology; D019453:Escherichia coli O157 / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D029968:Escherichia coli Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism*; D005517:Foodborne Diseases / Q000453:epidemiology / Q000382:microbiology; D017353:Gene Deletion; D015870:Gene Expression; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D008124:Locomotion; D010750:Phosphoproteins / Q000096:biosynthesis*; D020543:Proteome / Q000032:analysis; D018724:Spinacia oleracea / Q000382:microbiology; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014481:United States
chemical_list: D001639:Bicarbonates; D029968:Escherichia coli Proteins; C457340:LEE protein, E coli; D010750:Phosphoproteins; D020543:Proteome; C489568:RcsB protein, E coli; D014157:Transcription Factors