Open AccessCrystallization and preliminary X‐ray diffraction analysis of the barley yellow dwarf virus cap‐independent translation element
Author(s) -
Kraft Jelena J.,
Hoy Julie A.,
Miller W. Allen
Publication year - 2011
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309111007196
Subject(s) - barley yellow dwarf , crystallization , crystallography , monoclinic crystal system , messenger rna , biology , rna , transcription (linguistics) , microbiology and biotechnology , crystal structure , physics , chemistry , virus , genetics , gene , plant virus , linguistics , philosophy , thermodynamics
Barley yellow dwarf virus (BYDV) RNA lacks a 5′ m 7 GTP cap, yet it is translated efficiently because it contains a 105‐base BYDV‐like cap‐independent translation element (BTE) in the 3′ untranslated region (UTR). To understand how the BTE outcompetes the host mRNA for protein‐synthesis machinery, its three‐dimensional structure is being determined at high resolution. The purification using transcription from DNA containing 2′‐ O ‐methyl nucleotides and preliminary crystallographic analyses of the BTE RNA are presented here. After varying the BTE sequence and crystallization‐condition optimization, crystals were obtained that diffracted to below 5 Å resolution, with a complete data set being collected to 6.9 Å resolution. This crystal form indexes with an R merge of 0.094 in the monoclinic space group C 2, with unit‐cell parameters a = 316.6, b = 54.2, c = 114.5 Å, α = γ = 90, β = 105.1°.