Abstract
BackgroundEnterotoxigenic Escherichia coli (ETEC) is a major cause of infant and child mortality in developing countries. This enteric pathogen causes profuse watery diarrhea by elaborating one or more enterotoxins that intoxicate eukaryotic cells and ultimately leads to a loss of water to the intestinal lumen. Virulence is also dependent upon fimbrial adhesins that facilitate colonization of the small intestine.ResultsThe expression of CS1 fimbriae is positively regulated by Rns, a member of the AraC/XylS superfamily of transcriptional regulators. Based on fimbrial protein homology, CS1 fimbriae have been categorized as subclass 5b along with CS17, CS19, and PCFO71 fimbriae. In this study we show that Rns positively regulates the expression of these other subclass 5b members. DNase I footprinting revealed a Rns binding site adjacent to the -35 hexamer of each fimbrial promoter. The CS17 and PCFO71 fimbrial promoters carry a second Rns binding site centered at -109.5, relative to the Rns-dependent transcription start site. This second binding site is centered at -108.5 for the CS19 promoter. Mutagenesis of either site reduced Rns-dependent transcription from each promoter indicating that the molecules bound to these sites apparently function independently of one another, with each having an additive effect upon fimbrial promoter activation.ConclusionThis study demonstrates that the ETEC virulence regulator Rns is required for the expression of all known 5b fimbriae. Since Rns is also known to control the expression of additional ETEC fimbriae, including those within subclasses 5a and 5c, the inactivation or inhibition of Rns could be an effective strategy to prevent ETEC infections.
Highlights
Enterotoxigenic Escherichia coli (ETEC) is a major cause of infant and child mortality in developing countries
To determine if other subclass 5b fimbria are regulated by Rns, we cloned the promoters of CS17, CS19, and putative colonization factor O71 (PCFO71) into a promoterless Lac reporter plasmid that was integrated into the chromosomal attBHK022 site of K-12 strain MC4100 by site specific recombination
Quantitative enzymatic assays revealed that the reporter strains expressed 8 to 10 times more β-galactosidase when they were transformed with a Rns expression plasmid, pGPMRns, than when they were transformed with the rns::kan negative control plasmid pGPMRns2 (Table 1). When these results are combined with previous analyses of the coli surface antigen 1 (CS1) fimbrial promoter [15], they reveal that all known subclass 5b fimbrial promoters are activated by Rns
Summary
Enterotoxigenic Escherichia coli (ETEC) is a major cause of infant and child mortality in developing countries This enteric pathogen causes profuse watery diarrhea by elaborating one or more enterotoxins that intoxicate eukaryotic cells and leads to a loss of water to the intestinal lumen. Diarrheal disease can be caused by any one of several bacterial or viral pathogens, enterotoxigenic Escherichia coli (ETEC) is one of the most frequent causes of diarrhea in developing nations [3,4,5]. This pathogen causes profuse watery diarrhea by elaborating one or more enterotoxins. Pathogenicity is dependent upon the expression of fimbriae, which function as adherence fac-
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
