The metal- and DNA-binding activities of Helicobacter pylori NikR.
The pathogenic bacteria Helicobacter pylori require nickel as a cofactor of the enzymes urease and hydrogenase. One of the proteins that controls nickel homeostasis in this organism is Helicobacter pylori NikR (HpNikR), a homologue of nickel-dependent transcription factors from other organisms, which regulates the expression of multiple proteins such as the urease structural subunits and itself. To examine the properties of this protein, metal analysis was used to demonstrate that HpNikR can bind stoichiometric nickel or copper, and electronic absorption spectroscopy revealed that HpNikR binds nickel with picomolar affinity in what is likely a conserved square-planar site. In vitro DNA-binding assays revealed that HpNikR can bind directly to the promoter region of the ureA operon in response to nickel, and the location of the binding site was defined. Nickel also induces DNA binding to the nikR promoter sequence but the complex is much weaker. These experiments suggest that HpNikR directly controls the expression of multiple genes by binding to separate DNA sequences, and the possible mechanisms for differential regulation are discussed.
Abraham
Lihor O
LO
Department of Chemistry, University of Toronto, 80 St. George St., Toronto, Ont., Canada M5S 3H6.
Li
Yanjie
Y
Zamble
Deborah B
DB
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites; D004247:DNA / Q000378:metabolism*; D018983:DNA Footprinting; D017931:DNA Primers; D024202:Electrophoretic Mobility Shift Assay; D016480:Helicobacter pylori / Q000378:metabolism*; D008670:Metals / Q000378:metabolism*; D010957:Plasmids; D011401:Promoter Regions, Genetic
D001426:Bacterial Proteins; D017931:DNA Primers; D008670:Metals; D004247:DNA