Improving RNA Crystal Diffraction Quality by Postcrystallization Treatment.

Abstract

The crystallization and structural determination of large RNAs and their complexes remain major bottlenecks in the mechanistic analysis of cellular and viral RNAs. Here, we describe a protocol that combines postcrystallization dehydration and ion replacement that dramatically improved the diffraction quality of crystals of a large gene-regulatory tRNA-mRNA complex. Through this method, the resolution limit of X-ray data extended from 8.5 to 3.2 Å, enabling structure determination. Although this protocol was developed for a particular RNA complex, the general importance of solvent and counterions in nucleic acid structure may render it generally useful for crystallographic analysis of other RNAs.

• Authors: Jinwei Zhang, Adrian R Ferré-D'Amaré
• Journal: Methods in molecular biology (Clifton, N.J.) (Methods Mol Biol) Vol. 2323 Pg. 25-37 ( 2021) ISSN: 1940-6029 [Electronic] United States
• PMID: 34086271 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, N.I.H., Intramural, Research Support, U.S. Gov't, Non-P.H.S.)

Chemical References

• Ions
• RNA
• RNA, Transfer

Topics

• Crystallization (methods)
• Crystallography, X-Ray (methods)
• Desiccation
• Ions (chemistry)
• Nucleic Acid Conformation
• RNA (chemistry)
• RNA, Transfer (chemistry)