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Transcriptional activation of MerR family promoters in Cupriavidus metallidurans CH34

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Abstract

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 σ70–dependent promoters. In this study we report β-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.

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Acknowledgments

This work was supported by Biotechnology and Biological Sciences Research Council grant B10333 and a BBSRC studentship to DJJ. The Birmingham Functional Genomics laboratory was supported by the Biotechnology and Biological Sciences Research Council Joint Infrastructure Fund grant JIF13209.

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Authors and Affiliations

  1. School of Biosciences, University of Birmingham, Edgbaston, Birmingham, B15 2TT, UK

    Daniel J. Julian, Christopher J. Kershaw, Nigel L. Brown & Jon L. Hobman

  2. School of Biosciences, The University of Nottingham, Sutton Bonington Campus, Sutton Bonington, Loughborough, LE12 5RD, UK

    Jon L. Hobman

  3. College of Science and Engineering, The University of Edinburgh, Weir Building, The King’s Buildings, West Mains Road, Edinburgh, EH9 3JY, UK

    Nigel L. Brown

  4. Faculty of Life Sciences, The University of Manchester, Michael Smith Building, Oxford Road, Manchester, M13 9PT, UK

    Christopher J. Kershaw

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  1. Daniel J. Julian
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  4. Jon L. Hobman
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Correspondence to Jon L. Hobman.

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Julian, D.J., Kershaw, C.J., Brown, N.L. et al. Transcriptional activation of MerR family promoters in Cupriavidus metallidurans CH34. Antonie van Leeuwenhoek 96, 149–159 (2009). https://doi.org/10.1007/s10482-008-9293-4

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