2008
DOI: 10.1128/jb.00440-08
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Transient Shielding of Intimin and the Type III Secretion System of Enterohemorrhagic and Enteropathogenic Escherichia coli by a Group 4 Capsule

Abstract: Enterohemorrhagic and enteropathogenic Escherichia coli (EHEC and EPEC, respectively) strains represent a major global health problem. Their virulence is mediated by the concerted activity of an array of virulence factors including toxins, a type III protein secretion system (TTSS), pili, and others. We previously showed that EPEC O127 forms a group 4 capsule (G4C), and in this report we show that EHEC O157 also produces a G4C, whose assembly is dependent on the etp, etk, and wzy genes. We further show that at… Show more

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Cited by 41 publications
(43 citation statements)
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“…The cells were treated overnight with serum-free Dulbecco's modified Eagle's medium (DMEM) supplemented with mannose and HEPES and then infected with REPEC/pSR1 to monitor translocation. Bacterial cells grown overnight in LB broth were diluted 1:100 in Casamino Acids-DMEM (cDMEM) supplemented with 2.5 mM probenecid until they reached an optical density at 600 nm (OD 600 ) of 0.2 to 0.35. cDMEM is a tissue culture medium with low autofluorescence (28), and probenecid, a nonspecific anion inhibitor, facilitates CCF2-AM [coumarin-cephalosporin-fluorescein (2)-acetoxymethyl] loading by blocking its active efflux from cells (29). HeLa cells preloaded with CCF2-AM (GeneBLAzer In Vivo detection kit; Life Technologies, Carlsbad, CA) were infected with ϳ10 6 bacteria (multiplicity of infection [MOI] of 500 to 700).…”
Section: Methodsmentioning
confidence: 99%
“…The cells were treated overnight with serum-free Dulbecco's modified Eagle's medium (DMEM) supplemented with mannose and HEPES and then infected with REPEC/pSR1 to monitor translocation. Bacterial cells grown overnight in LB broth were diluted 1:100 in Casamino Acids-DMEM (cDMEM) supplemented with 2.5 mM probenecid until they reached an optical density at 600 nm (OD 600 ) of 0.2 to 0.35. cDMEM is a tissue culture medium with low autofluorescence (28), and probenecid, a nonspecific anion inhibitor, facilitates CCF2-AM [coumarin-cephalosporin-fluorescein (2)-acetoxymethyl] loading by blocking its active efflux from cells (29). HeLa cells preloaded with CCF2-AM (GeneBLAzer In Vivo detection kit; Life Technologies, Carlsbad, CA) were infected with ϳ10 6 bacteria (multiplicity of infection [MOI] of 500 to 700).…”
Section: Methodsmentioning
confidence: 99%
“…We have referred to this as the "22-min locus" (28). It has also been referred to as the gfc (group 4 capsule) locus (101). Recent studies have illustrated a role for this locus in group 4 CPS assembly (also Wzy dependent and essentially a variant of group 1) and in regulation of the heat shock response in E. coli (48,86).…”
Section: Structures and Phylogeny Of Opx Proteinsmentioning
confidence: 99%
“…It has been well documented that the master positive regulator of LEE genes, as well as of genes located outside the LEE, is the LEE-encoded regulator (Ler), whose main function is to counteract the repression exerted by H-NS on these genes (1,3,9,17,23,39,48,57,62,65,69). It has also been reported that Ler represses its own expression in a concentration-dependent fashion (4) and that it can also act as a negative regulator for other genes outside the LEE (1,61). The key role of Ler in LEE regulation makes its gene the main direct target of both positive and negative global regulators, such as H-NS, Hha, IHF, and QseA (9,20,59,60,69).…”
mentioning
confidence: 99%