RNA structure determination by solid-state NMR spectroscopy

A Marchanka, B Simon, G Althoff-Ospelt & T Carlomagno

Nature Communications


Knowledge of the RNA three-dimensional structure, either in isolation or as part of RNP complexes, is fundamental to understand the mechanism of numerous cellular processes. Because of its flexibility, RNA represents a challenge for crystallization, while the large size of cellular complexes brings solution-state NMR to its limits. Here, we demonstrate an alternative approach on the basis of solid-state NMR spectroscopy. We develop a suite of experiments and RNA labeling schemes and demonstrate for the first time that ssNMR can yield a RNA structure at high-resolution. This methodology allows structural analysis of segmentally labelled RNA stretches in high-molecular weight cellular machines-independent of their ability to crystallize- and opens the way to mechanistic studies of currently difficult-to-access RNA-protein assemblies.