A fast selenium derivatization strategy for crystallization and phasing of RNA structures Journal Article
| Authors: | Olieric, V.; Rieder, U.; Lang, K.; Serganov, A.; Schulze-Briese, C.; Micura, R.; Dumas, P.; Ennifar, E. |
| Article Title: | A fast selenium derivatization strategy for crystallization and phasing of RNA structures |
| Abstract: | 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′-OCH<sub>3</sub>-modified RNA sequences, prior to the replacement of 2′-OCH<sub>3</sub> groups with their 2′-SeCH<sub>3</sub> counterparts. The method exploits the similar crystallization properties of 2′-OCH<sub>3</sub>- and 2′-SeCH<sub>3</sub>-modified RNAs and has been successfully validated for two test cases. In addition, our data show that 2′-SeCH<sub>3</sub>-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. Copyright © 2009 RNA Society. |
| Keywords: | controlled study; rna; rna synthesis; base sequence; crystallography, x-ray; selenium; nucleic acid conformation; rna structure; crystallization; structure determination; x-ray crystallography; electron; radiolysis; rna sequence; hiv-1; staining and labeling |
| Journal Title: | RNA |
| Volume: | 15 |
| Issue: | 4 |
| ISSN: | 1355-8382 |
| Publisher: | Cold Spring Harbor Laboratory Press |
| Date Published: | 2009-04-01 |
| Start Page: | 707 |
| End Page: | 715 |
| Language: | English |
| DOI: | 10.1261/rna.1499309 |
| PUBMED: | 19228585 |
| PROVIDER: | scopus |
| PMCID: | PMC2661828 |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 5" - "Export Date: 30 November 2010" - "CODEN: RNARF" - "Source: Scopus" |
