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在2.9A分辨率下,T7RNA聚合酶延长复合物的结构

Tahir H Tahirov¹, Dmitry Temiakov, Michael Anikin

¹High Throughput Factory, RIKEN Harima Institute at SPring-8, 1-1-1 Kouto, Mikazuki-cho, Sayo, Hyogo 679-5148, Japan.

|November 8, 2002

概括

晶体结构显示,菌体T7RNA聚合酶在RNA-DNA杂交结合时经历了显著的域重组,从而实现了稳定的转录延长. 这为转录机制和与其他RNA聚合酶的相似性提供了洞察力.

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:

菌体T7RNA聚合酶 (T7RNAP) 是一种单个子单元的酶.
它执行转录类似于细菌和真核多子单元RNA聚合酶.
了解T7RNAP结构可以了解基本的转录机制.

研究的目的:

为了确定T7RNA聚合酶延长复合物的晶体结构.
阐明T7RNAP中转录延长的结构基础.
为了比较T7RNAP与多子单元RNA聚合酶的结构.

主要方法:

使用X射线结晶学来确定结构.
在存在8个基对RNA-DNA混合体的情况下分析了结构.

主要成果:

纳入RNA-DNA混合体诱导了氨基末端域的重大重新排列.
这涉及~130个残留的替代折叠和核心子域的重定向.
一个深厚的蛋白质腔形成,埋葬了RNA-DNA混合体,并创建了一个RNA退出途径.
下游DNA结合被静电相互作用稳定.

结论:

T7 RNAP结构解释了转录延长的机制.
在T7RNAP和多子单元RNA聚合酶之间存在结构上的相似之处.
这些发现为T7 RNAP功能和演变提供了结构基础.

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