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Article Dans Une Revue RNA Année : 2009

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

Résumé

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(3)-modified RNA sequences, prior to the replacement of 2'-OCH(3) groups with their 2'-SeCH(3) counterparts. The method exploits the similar crystallization properties of 2'-OCH(3)- and 2'-SeCH(3)-modified RNAs and has been successfully validated for two test cases. In addition, our data show that 2'-SeCH(3)-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.

Dates et versions

hal-00370924 , version 1 (25-03-2009)

Identifiants

Citer

Vincent Olieric, Ulrike Rieder, Kathrin Lang, Alexander Serganov, Clemens Schulze-Briese, et al.. A fast selenium derivatization strategy for crystallization and phasing of RNA structures.. RNA, 2009, 15 (4), pp.707-15. ⟨10.1261/rna.1499309⟩. ⟨hal-00370924⟩

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