Role of pH in Modulating RNA-Protein Interactions in TRBP2-dsRBD2: An Interplay between Conformational Dynamics and Electrostatic Interactions

Understanding RNA-protein interactions is crucial for uncovering the mechanisms of cellular processes and can provide insights into the basis of various diseases, paving the way for the development of targeted therapeutic interventions. Exposure to stress conditions, such as hypoxia, leads to a drop in intracellular pH, which, in turn, alters the ionization states of amino acid residues and RNA bases, affecting the charge distribution and electrostatic interactions between RNA and proteins. In addition, pH also perturbs the structure and dynamics of proteins via the disruption of H-bonds and ionic interactions. Thus, it is crucial to ascertain the role of pH in modulating such interactions. We have previously shown the role of conformational dynamics in the RNA-protein interaction in TAR RNA-binding protein (TRBP) double-stranded RNA-binding domains (dsRBD) 1 and 2 using solution-state NMR spectroscopy. The current study provides insights into the effect of pH on interactions between TRBP2-dsRBD2 and a dsRNA. Remarkably, it was observed that a unit decrease in pH leads to an increase in the flexibility of TRBP2-dsRBD2 in RNA-binding residues, as seen in NMR dynamics experiments, in addition to altering the charge distribution on the protein surface. This led us to propose a dynamics-driven model where the two effects of pH, electrostatic and conformational flexibility, counterbalance each other. Thus, it can be concluded that the overall binding affinity between the protein and RNA is governed by a delicate balance between its conformational dynamics and electrostatic interactions.