A fast selenium derivatization strategy for crystallization and phasing of RNA structures

Vincent Olieric, Ulrike Rieder, Kathrin Lang, Alexander Serganov, Clemens Schulze-Briese, Ronald Micura, Philippe Dumas, Eric Ennifar

Research output: Contribution to journalArticlepeer-review

45 Scopus citations

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′-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.

Original languageEnglish
Pages (from-to)707-715
Number of pages9
JournalRNA
Volume15
Issue number4
DOIs
StatePublished - Apr 2009
Externally publishedYes

Keywords

  • RNA structure
  • Selenium
  • Structure determination
  • X-ray crystallography

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