Transcriptional activation of MerR family promoters in Cupriavidus metallidurans CH34.
Metal responsive MerR family transcriptional regulators are widespread in bacteria and activate the transcription of genes involved in metal ion detoxification, efflux, or homeostasis, in response to the presence of cognate metal species in the cytoplasm. MerR family regulators recognize and bind to dyad symmetrical DNA sequences in specific promoters that have a spacer region between the -35 and -10 sequences which is longer than the canonical 16-18 bp spacer for other sigma(70)-dependent promoters. In this study we report beta-galactosidase assays of MerR family-regulated gene expression in the multiple metal resistant bacterium Cupriavidus metallidurans. A series of pMU2385 reporter plasmid derivatives containing cloned MerR family-activated promoters were used to determine metal ion-induced responses from different MerR family regulated promoters, as well as regulators cloned with the cognate promoter into pMU2385. Mercuric ion-responsive MerR and lead ion-responsive PbrR activity was confirmed using this assay system as well as MerR family activator activity on heterologous promoters PcopA, PcadA, and Pzcc from Escherichia coli, Pseudomonas aeruginosa and Bordetella pertussis, respectively. In C. metallidurans CH34, transcription from these promoters was activated by MerR family regulators encoded on the chromosome or megaplasmids in response to copper (PcopA), and lead (PcadA and PzccA), showing that MerR family activators in C. metallidurans can act on MerR family promoters from other organisms, which have sequence differences to the predicted C. metallidurans promoters.
Julian
Daniel J
DJ
School of Biosciences, University of Birmingham, Edgbaston, Birmingham, B15 2TT, UK.
Kershaw
Christopher J
CJ
Brown
Nigel L
NL
Hobman
Jon L
JL
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D049921:Cupriavidus / Q000187:drug effects / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D024881:Drug Resistance, Bacterial; D015964:Gene Expression Regulation, Bacterial; D007477:Ions / Q000494:pharmacology; D007854:Lead / Q000494:pharmacology; D008628:Mercury / Q000494:pharmacology; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics / Q000502:physiology*; D015533:Transcriptional Activation; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D007477:Ions; C051124:MerR protein, Bacteria; D007854:Lead; D001616:beta-Galactosidase; D008628:Mercury