Binding of the ERalpha nuclear receptor to DNA is coupled to proton uptake.
Nuclear receptors act as ligand-modulated transcription factors and orchestrate a plethora of cellular functions central to health and disease. Although studied for more than half a century, many mysteries surrounding the mechanism of action of nuclear receptors remain unresolved. Herein, using isothermal titration calorimetry (ITC) in conjunction with macromolecular modeling (MM), we provide evidence that the binding of the ERalpha nuclear receptor to its DNA response element is coupled to proton uptake by two ionizable residues, H196 and E203, located at the protein-DNA interface. Alanine substitution of these ionizable residues decouples protonation and hampers the binding of ERalpha to DNA by nearly 1 order of magnitude. Remarkably, H196 and E203 are predominantly conserved across approximately 50 members of the nuclear receptor family, implying that proton-coupled equilibrium may serve as a key regulatory switch for modulating protein-DNA interactions central to nuclear receptor function and regulation. Taken together, our findings unearth an unexpected but critical step in the molecular action of nuclear receptors and suggest that they may act as sensors of intracellular pH.
Deegan
Brian J
BJ
Department of Biochemistry and Molecular Biology and USylvester Braman Family Breast Cancer Institute, Leonard Miller School of Medicine, University of Miami, Miami, Florida 33136, USA.
Seldeen
Kenneth L
KL
McDonald
Caleb B
CB
Bhat
Vikas
V
Farooq
Amjad
A
D000595:Amino Acid Sequence; D002151:Calorimetry / Q000379:methods; D002467:Cell Nucleus / Q000378:metabolism*; D017124:Conserved Sequence; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D047628:Estrogen Receptor alpha / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D011522:Protons; D020218:Response Elements; D055672:Static Electricity; D013816:Thermodynamics
D047628:Estrogen Receptor alpha; D011522:Protons; D004247:DNA