Abstract
Nickel ions are crucial components for the catalysis of biological reactions in prokaryotic organisms. As an uncontrolled nickel trafficking is toxic for living organisms, nickel-dependent bacteria have developed tightly regulated strategies to maintain the correct intracellular metal ion quota. These mechanisms require transcriptional regulator proteins that respond to nickel concentration, activating or repressing the expression of specific proteins related to Ni(II) metabolism. In Streptomyces griseus, a Gram-positive bacterium used for antibiotic production, SgSrnR and SgSrnQ regulate the nickel-dependent antagonistic expression of two superoxide dismutase (SOD) enzymes, a Ni-SOD and a FeZn-SOD. According to a previously proposed model, SgSrnR and SgSrnQ form a protein complex in which SgSrnR works as repressor, binding directly to the promoter of the gene coding for FeZn-SOD, while SgSrnQ is the Ni(II)-dependent co-repressor. The present work focuses on the determination of the biophysical and functional properties of SgSrnR. The protein was heterologously expressed and purified from Escherichia coli. The structural and metal-binding analysis, carried out by circular dichroism, light scattering, fluorescence and isothermal titration calorimetry, showed that the protein is a well-structured homodimer, able to bind nickel with moderate affinity. DNase I footprinting and β-galactosidase gene reporter assays revealed that apo-SgSrnR is able to bind its DNA operator and activates a transcriptional response. The structural and functional properties of this protein are discussed relatively to its role as a Ni(II)-dependent sensor.
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Acknowledgements
The authors thank Prof. Stefano Ciurli for financial support and useful discussion. They also thank Prof. Paolo Neyroz for helpful assistance with fluorescence measurements and examination of the data. This work was supported by the Department of Pharmacy and Biotechnology of the University of Bologna through funds for fundamental research. YB and AZ are recipient of Ph.D. fellowships from the University of Bologna.
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Beniamino, Y., Pesce, G., Zannoni, A. et al. SrnR from Streptomyces griseus is a nickel-binding transcriptional activator. J Biol Inorg Chem 25, 187–198 (2020). https://doi.org/10.1007/s00775-019-01751-5
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DOI: https://doi.org/10.1007/s00775-019-01751-5