Crystallization and preliminary X-ray diffraction analysis of the barley yellow dwarf virus cap-independent translation element
Abstract
Barley yellow dwarf virus (BYDV) RNA lacks a 5' m^7GTP 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 C2, with unit-cell parameters a = 316.6, b = 54.2, c = 114.5 Å, α = γ = 90, β = 105.1°.
Additional Information
© 2011 International Union of Crystallography. Received 4 November 2010; Accepted 25 February 2011. This work was supported by NIH grant R01 GM067104 with an American Recovery and Reinvestment Act supplement. We thank Jay Nix at the Advanced Light Source of Lawrence Berkeley National Laboratory for assistance with data collection, Jennifer Doudna for advice, Mark Hargrove for laboratory support and Jen Kaiser for assistance with data processing.Attached Files
Published - xb5033.pdf
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Additional details
- PMCID
- PMC3087640
- Eprint ID
- 70213
- Resolver ID
- CaltechAUTHORS:20160908-152227723
- R01 GM067104
- NIH
- American Recovery and Reinvestment Act (ARRA)
- Created
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2016-09-09Created from EPrint's datestamp field
- Updated
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2021-11-11Created from EPrint's last_modified field