Mechanistic insights into the nickel-dependent allosteric response of the Helicobacter pylori NikR transcription factor

In Helicobacter pylori, the nickel-responsive NikR transcription factor plays a key role in regulating intracellular nickel concentrations, which is an essential process for survival of this pathogen in the acidic human stomach. Nickel binding to H. pylori NikR (HpNikR) allosterically activates DNA binding to target promoters encoding genes involved in nickel homeostasis and acid adaptation, to either activate or repress their transcription. We previously showed that HpNikR adopts an equilibrium between an open conformation and DNAbinding competent cis and trans states. Nickel binding slows down conformational exchange between these states and shifts the equilibrium toward the binding-competent states. The protein then becomes stabilized in a cis conformation upon binding the ureA promoter. Here, we investigate how nickel binding creates this response and how it is transmitted to the DNA-binding domains. Through mutagenesis, DNA-binding studies, and computational methods, the allosteric response to nickel was found to be propagated from the nickel-binding sites to the DNA-binding domains via the II-sheets of the metal-binding domain and a network of residues at the interdomain interface. Our computational results suggest that nickel binding increases protein rigidity to slow down the conformational exchange. A thymine base in the ureA promoter sequence, known to be critical for high affinity DNA binding by HpNikR, was also found to be important for the allosteric response, while a modified version of this promoter further highlighted the importance of the DNA sequence in modulating the response. Collectively, our results provide insights into regulation of a key protein for H. pylori survival.

Automated summary

The nickel-responsive NikR transcription factor in Helicobacter pylori plays a crucial role in regulating intracellular nickel concentrations and is essential for the survival of this pathogen in the human stomach. Nickel binding to H. pylori NikR allosterically activates DNA binding to target promoters involved in nickel homeostasis and acid adaptation. Through mutagenesis, DNA-binding studies, and computational methods, it was found that the allosteric response to nickel is propagated from the nickel-binding sites to the DNA-binding domains via specific residues and structural elements. These findings provide insights into the regulation of a key protein for H. pylori survival.