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(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.
authors.affiliation: Paul Scherrer Institute, Villigen, Switzerland.
authors.forename: Vincent
authors.initials: V
authors.lastname: Olieric
authors.forename: Ulrike
authors.initials: U
authors.lastname: Rieder
authors.forename: Kathrin
authors.initials: K
authors.lastname: Lang
authors.forename: Alexander
authors.initials: A
authors.lastname: Serganov
authors.forename: Clemens
authors.initials: C
authors.lastname: Schulze-Briese
authors.forename: Ronald
authors.initials: R
authors.lastname: Micura
authors.forename: Philippe
authors.initials: P
authors.lastname: Dumas
authors.forename: Eric
authors.initials: E
authors.lastname: Ennifar
mesh_terms: D001483:Base Sequence; D018360:Crystallography, X-Ray / Q000379:methods*; D015497:HIV-1 / Q000737:chemistry / Q000235:genetics; D009690:Nucleic Acid Conformation; D012313:RNA / Q000737:chemistry*; D012643:Selenium / Q000378:metabolism*; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D012313:RNA; D012643:Selenium