Histidine switch controlling pH-dependent protein folding and DNA binding in a transcription factor at the core of synthetic network devices.
Therapeutic strategies have been reported that depend on synthetic network devices in which a urate-sensing transcriptional regulator detects pathological levels of urate and triggers production or release of urate oxidase. The transcription factor involved, HucR, is a member of the multiple antibiotic resistance (MarR) protein family. We show that protonation of stacked histidine residues at the pivot point of long helices that form the scaffold of the dimer interface leads to reversible formation of a molten globule state and significantly attenuated DNA binding at physiological temperatures. We also show that binding of urate to symmetrical sites in each protein lobe is communicated via the dimer interface. This is the first demonstration of regulation of a MarR family transcription factor by pH-dependent interconversion between a molten globule and a compact folded state. Our data further suggest that HucR may be utilized in synthetic devices that depend on detection of pH changes.
Department of Biological Sciences, Louisiana State University, Baton Rouge, LA 70803, USA. agrove@lsu.edu.
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Deochand
Department of Biological Sciences, Louisiana State University, Baton Rouge, LA 70803, USA. agrove@lsu.edu.
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Perera
Department of Biological Sciences, Louisiana State University, Baton Rouge, LA 70803, USA. agrove@lsu.edu.
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Crochet
Department of Biological Sciences, Louisiana State University, Baton Rouge, LA 70803, USA. agrove@lsu.edu.
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Gilbert
Department of Biological Sciences, Louisiana State University, Baton Rouge, LA 70803, USA. agrove@lsu.edu.
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Newcomer
Department of Biological Sciences, Louisiana State University, Baton Rouge, LA 70803, USA. agrove@lsu.edu.
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Grove
D001665:Binding Sites; D004247:DNA / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006639:Histidine / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008024:Ligands; D008958:Models, Molecular; D008968:Molecular Conformation; D011485:Protein Binding; D017510:Protein Folding; D013816:Thermodynamics; D014157:Transcription Factors / Q000737:chemistry* / Q000378:metabolism*
D008024:Ligands; D014157:Transcription Factors; D006639:Histidine; D004247:DNA