Journal
RNA
Volume 15, Issue 4, Pages 707-715Publisher
COLD SPRING HARBOR LAB PRESS, PUBLICATIONS DEPT
DOI: 10.1261/rna.1499309
Keywords
X-ray crystallography; RNA structure; selenium; structure determination
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Funding
- Agence Nationale de Recherche sur le SIDA (ANRS)
- French Research Agency [PCV07-187047]
- Austrian Science Fund [P17864]
- Austrian Science Fund (FWF) [P17864] Funding Source: Austrian Science Fund (FWF)
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Site-specific 2'-methylseleno RNA labeling is a promising tool for tackling the phase problem in RNA crystallography. We have developed an efficient strategy for crystallization and structure determination of RNA and RNA/protein complexes based on preliminary crystallization screening of 2'-OCH3-modified RNA sequences, prior to the replacement of 2'-OCH3 groups with their 2'-SeCH3 counterparts. The method exploits the similar crystallization properties of 2'-OCH3- and 2'-SeCH3-modified RNAs and has been successfully validated for two test cases. In addition, our data show that 2'-SeCH3-modified RNA have an increased resistance to X-ray radiolysis in comparison with commonly used 5-halogen-modified RNA, which permits collection of experimental electron density maps of remarkable quality.
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