Genetics, Toxicity, and Distribution of Enterohemorrhagic Escherichia coli Hemolysin

The ability to produce enterohemolysin is regarded as a potential virulence factor for enterohemorrhagic Escherichia coli (EHEC) and is frequently associated with severe human diseases such as hemorrhagic colitis (HC) and the hemolytic uremic syndrome (HUS). The responsible toxin, which has also been termed EHEC-hemolysin (EHEC-Hly, syn. Ehx), belongs to the Repeats in Toxin (RTX)-family of pore-forming cytolysins and is characterized by the formation of incomplete turbid lysis zones on blood agar plates containing defibrinated sheep erythrocytes. Besides the expression of Shiga toxins (Stx) and the locus of enterocyte effacement (LEE), EHEC-Hly is a commonly used marker for the detection of potential pathogenic E. coli strains, although its exact role in pathogenesis is not completely understood. Based on the current knowledge of EHEC-Hly, this review describes the influence of various regulator proteins, explains the different mechanisms leading to damage of target cells, discusses the diagnostic role, and gives an insight of the prevalence and genetic evolution of the toxin.

Prevalence, Risk Factors, O Serogroups, and Virulence Profiles of Shiga Toxin–Producing Bacteria from Cattle Production Environments

Shiga toxin (Stx)–producing bacteria are important human pathogens that have been linked with cattle and associated food products. We recovered Stx-producing bacteria from 27.5% of cattle, 6.8% of water, and 2.3% of wildlife samples from a cattle production area during an 11-month period. Positive samples were found during every month and on 98% of sampling days. We recovered isolates from all cattle operations sampled, and prevalence within operations ranged from approximately 5 to 33%. Cattle prevalence was associated with the presence of Stx-producing bacteria in water and the production group and environment of cattle, with an interaction between production group and environment. Odds of recovering isolates from cattle were highest for groups of adult cows and their unweaned calves in pasture environments. Overall, 49 O serogroups were identified from 527 isolates. Seventy of the isolates contained virulence genes that encoded intimin and enterohemorrhagic Escherichia coli hemolysin. These were serogroups O111, O157, O109, O103, O145, O172, O84, O26, O108, O117, O126, O159, O5, O69, O74, O98, and O-rough. Our results suggest that the prevalence of Stx-producing bacteria can be relatively high in cattle, and associated factors may not be entirely similar to those reported for serotype O157:H7. Although Stx-producing bacteria were frequently detected, the strains may not be equally pathogenic for humans given the wide variety of serogroups and virulence genes. However, focusing on O157:H7 in food safety and surveillance programs may allow other Stx-producing bacteria, which appear to be widespread in cattle, to go undetected.

Regulation of Escherichia coli Hemolysin E Expression by H-NS and Salmonella SlyA

ABSTRACT The Escherichia coli hlyE gene (also known as clyA or sheA) codes for a novel pore-forming toxin. Previous work has shown that the global transcription factors FNR and CRP positively regulate hlyE expression by binding at the same site. Here in vivo transcription studies reveal that FNR occupies the hlyE promoter more frequently than CRP, providing a mechanism for the moderate upregulation of hlyE expression in response to two distinct environmental signals (oxygen and glucose starvation). It has been reported that H-NS interacts with two large regions of the hlyE promoter (PhlyE), one upstream of the −35 element and one downstream of the −10 element. Here we identify two high-affinity H-NS sites, H-NS I, located at the 3′ end of the extended upstream footprint, and H-NS II, located at the 5′ end of the extended downstream footprint. It is suggested that these high-affinity sites initiate the progressive formation of higher order complexes, allowing a range of H-NS-mediated regulatory effects at PhlyE. Finally, the identification of a SlyA binding site that overlaps the H-NS I site in PhlyE suggests a mechanism to explain how SlyA overproduction enhances hlyE expression by antagonizing the negative effects of H-NS.

Temperature- and H-NS-Dependent Regulation of a Plasmid-Encoded Virulence Operon Expressing Escherichia coli Hemolysin

ABSTRACT Proteins H-NS and Hha form a nucleoprotein complex that modulates expression of the thermoregulated hly operon of Escherichia coli. We have been able to identify two H-NS binding sites in the hly regulatory region. One of them partially overlaps the promoter region (site II), and the other is located about 2 kbp upstream (site I). In contrast, Hha protein did not show any preference for specific sequences. In vitro, temperature influences the affinity of H-NS for a DNA fragment containing both binding sites and H-NS-mediated repression of hly operon transcription. Deletion analysis of the hly regulatory region confirms the relevance of site I for thermoregulation of this operon. We present a model to explain the temperature-modulated repression of the hly operon, based on the experiments reported here and other, preexisting data.