Characterization of Mg²⁺ Distributions around RNA in Solution

Binding of metal ions is an important factor governing the folding and dynamics of RNA. Shielding of charges in the polyanionic backbone allows RNA to adopt a diverse range of folded structures that give rise to their many functions within the cell. Some RNA sequences fold only in the presence of Mg²⁺, which may be bound via direct interactions or occupy the more diffuse "ion atmosphere" around the RNA. To understand the driving forces for RNA folding, it is important to be able to fully characterize the distribution of metal ions around the RNA. In this work, a combined Grand Canonical Monte Carlo-Molecular Dynamics (GCMC-MD) method is applied to characterize Mg²⁺ distributions around folded RNA structures. The GCMC-MD approach identifies known inner- and outer-shell Mg²⁺ coordination, while also predicting new regions occupied by Mg²⁺ that are not observed in crystal structures but that may be relevant in solution, including the case of the Mg²⁺ riboswitch, for which alternate Mg²⁺ binding sites may have implications for its function. This work represents a significant step forward in establishing a structural and thermodynamic description of RNA-Mg²⁺ interactions and their role in RNA structure and function.