细菌RNA聚合酶促进体融化的早期中间体,通过时间解析的冷电子显微镜可视化
Ruth M Saecker1, Andreas U Mueller1, Brandon Malone1,2
1Laboratory of Molecular Biophysics, The Rockefeller University, New York, NY, USA.
Nature structural & molecular biology
|July 1, 2024
概括
细菌RNA聚合酶通过中间步骤形成转录复合体. 时间解析的冷EM可视化了在开放复合体形成期间的DNA开放和蛋白质重组.
科学领域:
- 分子生物学分子生物学
- 结构生物学 结构生物学
- 生物化学 生化学
背景情况:
- 细菌RNA聚合酶 (RNAPs) 通过短暂的中间体形成了对转录有竞争力的开放复合体 (RPo).
- 缺乏关于这些互转换的实时结构数据.
研究的目的:
- 在细菌RNAP促进剂开放复合形成过程中对中间体进行结构性表征.
- 阐明DNA开放和RPO组装的动态过程.
主要方法:
- 时间分辨率低温电子显微镜 (cryo-EM).
- 使用了大肠杆菌 σ70-RNAP 和 λPR促进体.
主要成果:
- 在120 ms和500 ms捕获了四种不同的中间值.
- 观察到快速的上游DNA泡脱配和逐步插入非模板链基.
- 记录了RNAP关闭和驱逐抑制 σ70 域的过程.
结论:
- 这项研究提供了对细菌RNAP开放复合体形成的首次实时结构洞察.
- 确定了关键的结构变化,包括DNA解和蛋白质域重组.
- 表明DNA序列影响RPO形成步骤,适用于许多细菌促进体.
相关概念视频
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