Induction of beta-lactamase production in Aeromonas hydrophila is responsive to beta-lactam-mediated changes in peptidoglycan composition.

We have studied the mechanism by which beta-lactam challenge leads to beta-lactamase induction in Aeromonas hydrophila through transposon-insertion mutagenesis. Disruption of the dd-carboxypeptidases/endopeptidases, penicillin-binding protein 4 or BlrY leads to elevated monomer-disaccharide-pentapeptide levels in A. hydrophila peptidoglycan and concomitant overproduction of beta-lactamase through activation of the BlrAB two-component regulatory system. During beta-lactam challenge, monomer-disaccharide-pentapeptide levels increase proportionately with beta-lactamase production and beta-lactamase induction is inhibited by vancomycin, which binds muro-pentapeptides. Taken together, these data strongly suggest that the Aeromonas spp. beta-lactamase regulatory sensor kinase, BlrB, responds to the concentration of monomer-disaccharide-pentapeptide in peptidoglycan.

Defining the growth conditions and promoter-proximal DNA sequences required for activation of gene expression by CreBC in Escherichia coli.

CreBC is a two-component system that controls the expression of a number of genes in Escherichia coli (called the cre regulon) that encode diverse functions, including intermediary metabolic enzymes. Using a reporter construct, we have shown that cre regulon gene expression is activated during growth in minimal media when glycolytic carbon sources are being fermented. It also is activated during aerobic growth when fermentation products are being used as carbon sources. CreB and CreC are essential for the activation of cre regulon gene expression, but CreA and CreD, encoded as part of the creABCD gene cluster, are not. CreB binds to a TTCACnnnnnnTTCAC direct repeat (the cre tag) in vitro, and this sequence, which is associated with cre regulon gene promoters, is required for the control of gene expression in vivo. These observations support the hypothesis that CreBC is a functional two-component system involved in the metabolic control of transcription in E. coli and confirm that CreB is a DNA binding transcriptional regulator.