Crystallization and preliminary crystallographic analysis of T7 RNA polymerase elongation complex

被引:12
作者
Temiakov, D
Tahirov, TH
Anikin, M
McAllister, WT
Vassylyev, DG
Yokoyama, S
机构
[1] SUNY Hlth Sci Ctr, Dept Microbiol & Immunol, Morse Inst Mol Genet, Brooklyn, NY 11203 USA
[2] RIKEN, Harima Inst SPring 8, High Throughput Factory, Sayo, Hyogo 6795148, Japan
[3] RIKEN, Harima Inst SPring 8, Cellular Signaling Lab, Sayo, Hyogo 6795148, Japan
[4] RIKEN, Harima Inst SPring 8, Structurome Project, Sayo, Hyogo 6795148, Japan
[5] Univ Tokyo, Grad Sch Sci, Dept Biochem & Biophys, Bunkyo Ku, Tokyo 1130033, Japan
[6] RIKEN, Genom Sci Ctr, Yokohama, Kanagawa 2300045, Japan
来源
ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY | 2003年 / 59卷
关键词
D O I
10.1107/S0907444902019777
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Stable transcription-elongation complexes consisting of T7 RNA polymerase (molecular mass 99 kDa) in association with a nucleic acid scaffold consisting of an 8 bp RNA-DNA hybrid and 10 bp of downstream DNA were assembled and crystallized by the sitting-drop vapour-diffusion technique under near-physiological conditions. The crystals diffract beyond 2.6 Angstrom resolution and belong to space group P1, with unit-cell parameters a = 79.91, b = 84.97, c = 202 Angstrom, alpha = 90.36, beta = 92.97, gamma = 109.94degrees. An unambiguous molecular-replacement solution was found using the C-terminal portion of the T7 RNA polymerase structure from the early initiation complex as a search model. Model building and structure refinement are now in progress.
引用
收藏
页码:185 / 187
页数:3
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