在T7RNA聚合酶中转位和酶活性的结构机制
Y Whitney Yin1, Thomas A Steitz
1Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06520, USA.
Cell
|March 16, 2004
概括
RNA聚合酶作为一个分子电机. 它的构造变化是由酸盐释放驱动的,在转录过程中使DNA转位和链分离成为可能.
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 生物化学
背景情况:
- RNA聚合酶是转录所必需的分子电机.
- 了解它的机制需要详细的结构洞察到不同的功能状态.
研究的目的:
- 阐明RNA聚合酶在核酸添加和转位过程中的功能结构基础.
- 研究酶构成在DNA链分离和转位中的作用.
主要方法:
- 使用X射线晶体学来确定菌体T7RNA聚合酶的结构.
- 获得了延伸阶段基质和转移前产物复合物的结构.
主要成果:
- 在核酸添加周期期间,确定了两个不同的酶构造.
- 一个五螺旋子域采用不同的定向,由输入的核酸三酸盐 (NTP) 或产品酸盐 (PPi) 稳定.
- 缺少NTP或PPi可以稳定与转位和DNA链分离相关的构造.
结论:
- 产品PPi解离会触发RNA聚合酶的结构变化.
- 这种形状变化对于实现转位和DNA链分离至关重要.
- RNA聚合酶利用不同的结构状态将化学能量与转录中的机械工作结合起来.
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