PUSHING THE FRONTIERS OF RNA STRUCTURAL DYNAMICS USING SITE-SPECIFIC LABELING FOR SOLUTION NMR SPECTROSCOPY

Abstract

The roles of RNAs (ribonucleic acids) in various cellular processes are underpinned by the unique architectures and range of dynamic motions they adopt. Subtle adjustments to RNA composition, structure, or even environment may confer profound changes to their stability and functions. Solution Nuclear Magnetic Resonance (NMR) Spectroscopy offers high-resolution and non-destructive approaches to explore these motions and structures, thereby enhancing our understanding of these biomolecules. However, the current median size of RNA structures probed by solution NMR spectroscopy is approximately 25 nucleotides (nts). The sparse distribution of high molecular weight RNA structures and dynamics data from NMR spectroscopy is ascribed to the lack of chemical shift diversity and line broadening. Here, the fundamental bases for line broadening effects are explored, accompanied by huge strides in the field to combat them. Recently, we have synthesized some site-specific isotopic labels that have demonstrated superior spectroscopic properties, thus, making them applicable to large RNAs. We showcase how the application of these isotopic labels expands our toolbox to study various RNA ranging in size from 27 nts to 232 nts by NMR spectroscopy.