A Solid View on RNA: Solid-State NMR of RNA and RNP Complexes

Abstract

To date, substantial progresses have been made in the structure determination of membrane proteins and amyloid fibrils by solid-state NMR, while significantly fewer studies have addressed the structure of RNA or protein-RNA complexes (RNP). Nevertheless, the application of ssNMR to study large RNP complexes holds excellent promises, due to the independence of the ssNMR line widths from the molecular size. Here, I will present the ssNMR-based structure of the complex consisting of the the 26mer box C/D RNA and the protein L7Ae, together with the experimental strategy that we used to obtain it. This includes experiments for the assignment of RNA resonances and collection of structural restraints, as well as experiments to determine the protein-RNA contacts. In addition I will discuss the application of proton-detection in ssNMR of RNA. Finally, I will show first ssNMR spectra of a RNA in the context of the 400 kDa box C/D RNP.