Evaluation of the regulatory model for Ni 2+ sensing by Nur from Streptomyces coelicolor

Streptomyces coelicolor is a soil-dwelling bacterium that is medically important due to its ability to produce several antibiotics, and nickel accumulation within this organism has been shown to prevent the production of the antibiotic undecylprodigiosin. The transcriptional repressor important in r...

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Bibliographic Details
Published in:Journal of inorganic biochemistry 2020-02, Vol.203, p.110859
Main Authors: Manley, Olivia M, Myers, Paisley D, Toney, Denise J, Bolling, Katherine F, Rhodes, Lauren C, Gasparik, Jessica L, Grossoehme, Nicholas E
Format: Article
Language:English
Subjects:
Bacterial Proteins - chemistry
Bacterial Proteins - metabolism
Binding Sites
DNA - metabolism
Nickel - metabolism
Protein Binding
Repressor Proteins - chemistry
Repressor Proteins - metabolism
Streptomyces coelicolor - chemistry
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Summary:Streptomyces coelicolor is a soil-dwelling bacterium that is medically important due to its ability to produce several antibiotics, and nickel accumulation within this organism has been shown to prevent the production of the antibiotic undecylprodigiosin. The transcriptional repressor important in regulation of nickel uptake is the homodimeric Nur, a member of the Fur family. Nur contains two metal-binding sites per monomer: the M-site and the Ni-site. The work described here seeks to determine the roles of each of the metal-binding sites to establish a model of Nur activity through mutational studies, metal titrations, and fluorescence anisotropy. Through these studies, a model of Nur activity is proposed in which femtomolar metal binding to one M-site of Nur prompts DNA-binding, and metal binding to the second M-site fully activates the protein. Evidence is provided that shows cooperative metal binding to the Ni-site, but this process dampens affinity for promoter DNA.
ISSN:1873-3344