Solid-state NMR of RNA

We have pioneered the application of solid-state NMR (ssNMR) to RNA and RNPs. ssNMR is particularly suitable for RNA as part of large complexes, because the quality of ssNMR spectra is independent of the size of the molecule. In addition, ssNMR does not require crystals, and the sample preparation process preserves a high level of hydration, which is essential to support proper native folding of RNA.

We have determined the first de novo high-resolution structure of RNA by ssNMR (Marchanka et al., Angewandte Chemie, 2013; Marchanka et al., Nature Communications, 2015; Figure 1) and have developed methods for proton-detected ssNMR of RNA and identification of base-pairing patterns (Marchanka et al., Chemical Communications, 2018). Furthermore, we have determined the first NMR structure of an RNA–protein complex using only ssNMR-derived data. We believe that there remains significant unexplored potential for ssNMR of RNA and RNPs, which we are exploiting for both methodological developments and novel areas of application.

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Figure 1: solid-state NMR of box C/D kink-turn RNA
Figure 1. Left: ribose region of 2D 13C,15N-TEDOR-13C,13C-PDSD spectra of the Pf Box C/D RNA in complex with L7Ae. (a) G,U-labelled-RNA; (b) A,U-labelled-RNA. Intra- and inter-nucleotide correlations are labelled in green and red, respectively. Selected sequential correlations are indicated with dashed lines. Right: overlay of the 10 lowest-energy structures of the Pf Box C/D RNA in complex with L7Ae, determined from ssNMR data.