EXPANDING THE TOOLKIT: STRUCTURE, DYNAMICS, AND DRUG INTERACTIONS OF THE “PRIMING LOOP” FROM HEPATITIS B VIRUS PRE-GENOMIC RNA BY SOLUTION NMR SPECTROSCOPY

Abstract

RNAs are dynamic macromolecules that function as essential components of biological pathways that result in human disease, making them attractive therapeutic targets. Yet, RNA structural biology lags significantly behind that of proteins, limiting mechanistic understanding of RNA chemical biology. Fortunately, solution NMR spectroscopy can probe the structure, dynamics, and interactions of RNA in solution at atomic resolution, opening the door to their functional understanding. However, NMR analysis of RNA – with only four unique ribonucleotide building blocks – suffers from spectral crowding and broad linewidths, especially as RNAs grow in size. One effective strategy to overcome these challenges is to introduce NMR-active stable isotopes into RNA in an atom- and position-specific manner. Here, we outline the development of labeling technologies, their use in benefiting RNA dynamics measurements, and applications to study the structure, dynamics, and interactions of a conserved regulatory RNA stem-loop from hepatitis B virus that is critical for viral replication.