Protein–RNA footprinting: an evolving tool

AbstractAs more RNA molecules with important cellular functions are discovered, there is a strong need to characterize their structures, functions, and interactions. Chemical and enzymatic footprinting methods are used to map RNA secondary and tertiary structure, to monitor ligand interactions and conformational changes, and in the study of protein–RNA interactions. These methods provide data at single‐nucleotide resolution that nicely complements the structural information available from X‐ray diffraction, nuclear magnetic resonance spectroscopy (NMR), or cryo‐electron microscopy. Footprinting methods also complement the dynamic information derived from single‐molecule Förster resonance energy transfer. RNA footprinting tools have been used for decades, but we have recently seen spectacular advances, for instance, the use in combination with massive parallel sequencing techniques. Large libraries of RNA molecules (small or large in size) can now be probed in high‐throughput manner when RNA footprinting methods are combined with fluorescent probe technologies and automation. In this article, after a brief historical overview, we summarize recent advances in RNA–protein footprinting methodologies that now integrate tools for massive parallel analysis. WIREs RNA 2011, 3:557–566. doi: 10.1002/wrna.1119This article is categorized under: RNA Interactions with Proteins and Other Molecules > Protein–RNA Recognition RNA Interactions with Proteins and Other Molecules > RNA–Protein Complexes RNA Methods > RNA Analyses In Vitro and In Silico