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 88 bp 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 β-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 88 bp region upstream of the ehx operon contains a cis-acting element to which H-NS binds and negatively regulates expression of enterohemolysin.