title: The role for TolA in enterohemorrhagic Escherichia coli pathogenesis and virulence gene transcription.
abstract: Loss of the periplasm spanning protein TolA in Escherichia coli leads to activation of the Rcs phosphorelay, and is required for full virulence in Gram-negative pathogens such as Salmonella enterica and Dickeya dadantii. This study explores the role for TolA in the pathogenesis of enterohemorrhagic E. coli (EHEC) and the effect of its mutation on the transcription of key EHEC virulence genes controlled by Rcs phosphorelay, including the type III secretion system (T3SS) (espA and tir), the E. coli common pilus (ecpA), and motility (fliC). Promoter activity for T3SS regulator ler was substantially higher following inactivation of tolA, and corresponded with a similar elevation in espA and tir transcription. Likewise, ecpA transcription was increased in EHECΔtolA. Conversely, and in-line with previous studies, inactivation of tolA resulted in complete loss of motility and decreased fliC transcription. For all genes examined, altered transcription observed for EHECΔtolA was dependent on the outer-membrane lipoprotein RcsF. Despite elevated virulence gene transcription, in tolA deleted strains virulence of EHEC in the Galleria mellonella wax worm model was substantially attenuated in a manner at least partly dependent on RcsF, and adherence to cultured HT-29 colonic epithelial cells was markedly reduced. The results of this study broaden the role for TolA in EHEC pathogenesis, and suggest that significant outer-membrane perturbations are able to promote transcription of important EHEC adherence factors.
authors.affiliation: University of South Florida, Department of Cell Biology, Microbiology and Molecular Biology, Tampa, FL 33620, USA. Electronic address: jkmorgan@mail.usf.edu.
authors.forename: Jason K
authors.initials: JK
authors.lastname: Morgan
authors.affiliation: University of South Florida, Department of Cell Biology, Microbiology and Molecular Biology, Tampa, FL 33620, USA.
authors.forename: Jose A
authors.initials: JA
authors.lastname: Ortiz
authors.affiliation: University of South Florida, Department of Cell Biology, Microbiology and Molecular Biology, Tampa, FL 33620, USA.
authors.forename: James T
authors.initials: JT
authors.lastname: Riordan
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D054324:Enterohemorrhagic Escherichia coli / Q000235:genetics* / Q000472:pathogenicity / Q000502:physiology; D004927:Escherichia coli Infections / Q000382:microbiology; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D055786:Gene Knockout Techniques; D007915:Lepidoptera; D008124:Locomotion; D014158:Transcription, Genetic; D014774:Virulence; D037521:Virulence Factors / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D029968:Escherichia coli Proteins; D037521:Virulence Factors; C078611:rcsF protein, E coli; C062093:tolA protein, E coli
keywords: Adherence genes; EHEC; Outer-membrane integrity; Rcs phosphorelay; RcsF; TolA