Aeromonas hydrophila AmpH and CepH β-lactamases: derepressed expression in mutants of Escherichia coli lacking creB

• Published in: Journal of antimicrobial chemotherapy 2000-11, Vol.46 (5), p.695-702
• Main Authors: Avison, Matthew B., Niumsup, Pannika, Walsh, Timothy R., Bennett, Peter M.
• Format: Article
• Language: English
• Subjects: Aeromonas hydrophila; Aeromonas hydrophila - enzymology; Aeromonas hydrophila - genetics; Amino Acid Sequence; ampH gene; AmpH protein; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Bacteriology; Base Sequence; beta-Lactamases - genetics; beta-Lactamases - metabolism; Biological and medical sciences; Carrier Proteins - genetics; Carrier Proteins - metabolism; cepH gene; CepH protein; Cephalosporinase - genetics; Cephalosporinase - metabolism; creB gene; Cyclic AMP Response Element-Binding Protein - genetics; Cyclic AMP Response Element-Binding Protein - physiology; Escherichia coli; Escherichia coli - enzymology; Escherichia coli - genetics; Escherichia coli Proteins; Fundamental and applied biological sciences. Psychology; Humans; Metabolism. Enzymes; Microbiology; Molecular Sequence Data; Mutation - genetics; Penicillin-Binding Proteins; Sequence Analysis, DNA - methods
• ISSN: 0305-7453; 1460-2091; 1460-2091
• DOI: 10.1093/jac/46.5.695

The class 1 cephalosporinase (CepH) and class 2d oxacillinase (AmpH) from an Aeromonas hydrophila clinical isolate, strain T429125, have been cloned and sequenced. Both enzymes are typical of their equivalents in other species of Aeromonas. Both cloned β-lactamase genes were expressed at a low level in a standard laboratory Escherichia coli strain, but when cloned into a cre deletion E. coli mutant, they were expressed at significantly higher levels. Specific disruption of the creB gene resulted in similar increased levels of β-lactamase expression, so it was concluded that CreB represses the transcription of ampH and cepH in a cre+ E. coli strain. The expression of cepH was four times that of ampH in the δcre mutant because of an additional factor encoded on the cloned T429125 chromosomal fragment containing cepH. This factor was able to trans-activate expression of co-resident ampH in the δcre mutant such that expression of the two genes was approximately equal. The entire cepH-containing fragment was sequenced, but it contained no genes that were obviously related to any known class of DNA-binding protein.