使用EF-Tu•GTP进行延长性核糖体的冷-EM,阐明tRNA校对
Anna B Loveland1, Gabriel Demo1,2, Andrei A Korostelev3
1RNA Therapeutics Institute, Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA, USA.
Nature
|July 3, 2020
概括
核糖体使用动态解码过程校对转移RNA (tRNA). 通过延长因子EF-Tu的GTP水解释放tRNA,使核糖体能够锁定正确的tRNA或拒绝不正确的.
科学领域:
- 分子生物学
- 结构生物学
- 生物化学
背景情况:
- 核糖体通过校对由延长因子EF-Tu传递的氨基酸tRNA来确保翻译的准确性.
- 想象GTP催化延长和核糖体的校对机制是一个挑战.
研究的目的:
- 在GTP催化延长过程中阐明核糖体tRNA校对的分子机制.
- 在氨基酸tRNA选择过程中可视化核糖体的动态状态.
主要方法:
- 时间分辨率低温电子显微镜 (cryo-EM).
- 在EF-Tu•GTP输送氨基酸tRNA后分析了33种不同的核糖体状态.
主要成果:
- 核糖体解码中心在GTP水解前后动态监测子-抗子相互作用.
- 通过EF-Tu的GTP水解释放tRNA,使30S子单元能够锁定相关tRNA.
- 在初始选择和校对过程中,近同类tRNA不被锁定并被解离.
结论:
- 核糖体校对包括GTP水解后的动态监测和构造变化.
- 核糖体通过一种涉及初始选择和水解后校对的机制有效地排斥不正确的tRNA.
- 在初始选择和校对状态之间存在结构上的相似性,确保精确的蛋白质合成.
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