A fast selenium derivatization strategy for crystallization and phasing of RNA structures.
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.
Olieric
Vincent
V
Paul Scherrer Institute, Villigen, Switzerland.
Rieder
Ulrike
U
Lang
Kathrin
K
Serganov
Alexander
A
Schulze-Briese
Clemens
C
Micura
Ronald
R
Dumas
Philippe
P
Ennifar
Eric
E
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*
D012313:RNA; D012643:Selenium