pH-responsive DNA-binding activity of Helicobacter pylori NikR.
Helicobacter pylori NikR (HpNikR) is a nickel-responsive transcription factor. In addition to a role in nickel homeostasis, HpNikR is proposed to serve as a master activator-repressor for H. pylori acid adaptation by directly or indirectly regulating the expression of a battery of genes. One potential mechanism of this regulation is modulation of the DNA-binding activity of HpNikR by the decrease in internal pH that occurs upon exposure to acidic shock. To test this hypothesis, several properties of HpNikR were investigated under acidic conditions. At pH 5.8, the secondary and quaternary structures of the protein are not affected, and it still binds stoichiometric nickel in the same site, although with a slightly weaker affinity than that at pH 7.6. DNA-binding assays performed at pH 5.8 reveal that, in contrast to pH 7.6, HpNikR binds to the ureA promoter in a nickel-independent fashion. Binding to the nikR promoter at the lower pH is nickel dependent, however. Deletion of amino acids 3-11 of HpNikR abolished the nickel-responsive activity and enhanced nonspecific DNA binding. Site-directed mutagenesis of HpNikR indicates that either Asp7 or Asp8 in the N-terminus of HpNikR plays a part in the activation of DNA binding. Furthermore, Lys6 contributes selectively to complex formation with the nikR promoter sequence. The direct influence of pH on the activity of HpNikR may be critical to the role of this activator-repressor in the viability of H. pylori.
Li
Yanjie
Y
Department of Chemistry, University of Toronto, Toronto, Ontario, Canada M5S 3H6.
Zamble
Deborah B
DB
D020816:Amino Acid Motifs; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004269:DNA, Bacterial / Q000737:chemistry / Q000378:metabolism*; D016480:Helicobacter pylori / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D009532:Nickel / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D011487:Protein Conformation; D012097:Repressor Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D004269:DNA, Bacterial; C523724:NikR protein, Helicobacter pylori; D012097:Repressor Proteins; D009532:Nickel