Determining RNA three-dimensional structures using low-resolution data

doi:10.1016/j.jsb.2011.12.024

Determining RNA three-dimensional structures using low-resolution data

2012-09-01

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DOI：10.1016/j.jsb.2011.12.024

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Knowing the 3-D structure of an RNA is fundamental to understand its biological function. Nowadays X-ray crystallography and NMR spectroscopy are systematically applied to newly discovered RNAs. However, the application of these high-resolution techniques is not always possible, and thus scientists must turn to lower resolution alternatives. Here, we introduce a pipeline to systematically generate atomic resolution 3-D structures that are consistent with low-resolution data sets. We compare and evaluate the discriminative power of a number of low-resolution experimental techniques to reproduce the structure of the Escherichia colt tRNA(VAL) and P4-P6 domain of the Tetrahymena thermophila group I intron. We test single and combinations of the most accessible low-resolution techniques, i.e. hydroxyl radical footprinting (OH), methidiumpropyl-EDTA (MPE), multiplexed hydroxyl radical cleavage (MOHCA), and small-angle X-ray scattering (SAXS). We show that OH-derived constraints are accurate to discriminate structures at the atomic level, whereas EDTA-based constraints apply to global shape determination. We provide a guide for choosing which experimental techniques or combination of thereof is best in which context. The pipeline represents an important step towards high-throughput low-resolution RNA structure determination. (C) 2012 Elsevier Inc. All rights reserved.

Keywords:

RNA

3-D

Structure

Low-resolution

Footprinting

MPE

MOHCA

SAXS

MC-Fold

MC-Sym

Determining RNA three-dimensional structures using low-resolution data

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