Up-regulation of both intimin and eae-independent adherence of shiga toxigenic Escherichia coli O157 by ler and phenotypic impact of a naturally occurring ler mutation.
Shiga toxigenic Escherichia coli (STEC) strains are important human pathogens which are capable of causing diarrhea, hemorrhagic colitis, and the potentially fatal hemolytic-uremic syndrome (HUS). An important virulence trait of certain STEC strains, such as those belonging to serogroup O157, is the capacity to produce attaching and effacing (A/E) lesions on enterocytes, a property encoded by the locus for enterocyte effacement (LEE). LEE contains the eae gene, which encodes intimin, an outer membrane protein which mediates the intimate attachment of bacteria to the host epithelial cell surface, and eae is routinely used as a marker for LEE-positive STEC strains. However, the O157:H(-) STEC strain 95SF2 carries eae but did not produce A/E lesions on HEp-2 cells, as judged by a fluorescent actin staining assay. In this assay, 95SF2 adhered poorly to the HEp-2 cells, and those that did bind exhibited abnormal cell division. In contrast, the O157:H7 STEC strain EDL933 adhered strongly and produced typical A/E lesions. We have demonstrated that 95SF2 carries a defective LEE regulatory gene, ler, with a single base change with respect to that published for ler of EDL933, resulting in an Ile(57)-to-Thr substitution. Ler shows homology to H-NS-like regulators, which are modulators of transcription, and the mutation occurs in a domain implicated in oligomerization. 95SF2 was able to adhere and produce A/E lesions on HEp-2 cells when EDL933 ler was expressed from a multicopy plasmid. Conversely, introduction of a plasmid carrying 95SF2 ler into EDL933 abolished adherence and capacity to form A/E lesions. Studies with eae deletion derivatives of 95SF2 and EDL933 demonstrated that the ler-mediated adherence to HEp-2 cells is largely independent of intimin. We have also demonstrated that EDL933 ler, but not 95SF2 ler, increases the level of intimin in O157 STEC.
Ogierman
M A
MA
Molecular Microbiology Unit, Women's and Children's Hospital, North Adelaide, South Australia 5006, Australia.
Paton
A W
AW
Paton
J C
JC
D018829:Adhesins, Bacterial; D000595:Amino Acid Sequence; D001422:Bacterial Adhesion; D001425:Bacterial Outer Membrane Proteins / Q000502:physiology*; D001427:Bacterial Toxins / Q000633:toxicity*; D002352:Carrier Proteins; D019453:Escherichia coli O157 / Q000235:genetics / Q000502:physiology*; D029968:Escherichia coli Proteins; D005798:Genes, Bacterial; D006801:Humans; D008969:Molecular Sequence Data; D010641:Phenotype; D022603:Shiga Toxins; D015854:Up-Regulation
D018829:Adhesins, Bacterial; D001425:Bacterial Outer Membrane Proteins; D001427:Bacterial Toxins; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D022603:Shiga Toxins; C073819:eaeA protein, E coli