Histone-like nucleoid-structuring protein represses transcription of the ehx operon carried by locus of enterocyte effacement-negative Shiga toxin-expressing Escherichia coli.
Shiga toxin-producing Escherichia coli (STEC) are a significant cause of zoonotic foodborne diarrheal disease in industrialized nations. In addition to Shiga toxin most STEC produce the enterohemolysin (EhxA) toxin. The EhxA toxin is encoded by the ehxCABD operon located on the large plasmid carried by STEC, yet its role in pathogenesis is unknown. A histone-like nucleoid-structuring protein (H-NS) null mutant of STEC O91:H21 strain B2F1 displayed a hyper-hemolytic phenotype, was defective in binding to human colonic epithelial cells, and was non-motile. We concluded that H-NS modulated expression of several genes in B2F1 including the ehx operon. Electrophoretic mobility shift assays indicate that H-NS binds to an 88bp region of DNA upstream of the ehxC start codon. To determine if the same region of DNA was sensitive to repression by H-NS, a transcriptional fusion was constructed between the putative promoter region of ehx and a promoterless lacZ gene. The beta-galactosidase activity detected was low in E. coli that produced H-NS but was significantly higher in the H-NS null background. Taken together, the data indicates that in STEC the 88bp region upstream of the ehx operon contains a cis-acting element to which H-NS binds and negatively regulates expression of enterohemolysin.
Rogers
Miles T
MT
Department of Biological Sciences, Western Michigan University, 1903W. Michigan Ave., Kalamazoo, MI 49071, USA.
Zimmerman
Rachel
R
Scott
Maria E
ME
D019677:Artificial Gene Fusion; D001422:Bacterial Adhesion; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004269:DNA, Bacterial / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial; D017930:Genes, Reporter; D006461:Hemolysis; D006801:Humans; D008124:Locomotion; D009876:Operon; D010750:Phosphoproteins / Q000235:genetics; D011485:Protein Binding; D054323:Shiga-Toxigenic Escherichia coli / Q000472:pathogenicity / Q000502:physiology*; D014158:Transcription, Genetic; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; C031908:H-NS protein, bacteria; C457340:LEE protein, E coli; D010750:Phosphoproteins; D001616:beta-Galactosidase