RNA聚合酶上游面与β'亚基钳域之间的调控通讯。
Regulated communication between the upstream face of RNA polymerase and the beta' subunit jaw domain.
作者信息
Wigneshweraraj Siva R, Burrows Patricia C, Nechaev Sergei, Zenkin Nikolay, Severinov Konstantin, Buck Martin
机构信息
Department of Biological Sciences, Imperial College London, London SW7 2AZ, UK.
出版信息
EMBO J. 2004 Oct 27;23(21):4264-74. doi: 10.1038/sj.emboj.7600407. Epub 2004 Oct 7.
Abstract
We used bacteriophage T7-encoded transcription inhibitor gene protein 2 (gp2) as a probe to study the contribution of the Escherichia coli RNA polymerase (RNAP) beta' subunit jaw domain--the site of gp2 binding--to activator and ATP hydrolysis-dependent open complex formation by the sigma(54)-RNAP. We show that, unlike sigma(70)-dependent transcription, activated transcription by sigma(54)-RNAP is resistant to gp2. In contrast, activator and ATP hydrolysis-independent transcription by sigma(54)-RNAP is highly sensitive to gp2. We provide evidence that an activator- and ATP hydrolysis-dependent conformational change involving the beta' jaw domain and promoter DNA is the basis for gp2-resistant transcription by sigma(54)-RNAP. Our results establish that accessory factors bound to the upstream face of the RNAP, communicate with the beta' jaw domain, and that such communication is subjected to regulation.
摘要
我们使用噬菌体T7编码的转录抑制基因蛋白2（gp2）作为探针，来研究大肠杆菌RNA聚合酶（RNAP）β'亚基颌结构域（gp2的结合位点）对σ⁵⁴-RNAP激活剂和ATP水解依赖性开放复合物形成的贡献。我们发现，与依赖σ⁷⁰的转录不同，σ⁵⁴-RNAP的激活转录对gp2具有抗性。相反，σ⁵⁴-RNAP的激活剂和ATP水解非依赖性转录对gp2高度敏感。我们提供的证据表明，涉及β'颌结构域和启动子DNA的激活剂和ATP水解依赖性构象变化是σ⁵⁴-RNAP对gp2抗性转录的基础。我们的结果表明，与RNAP上游面结合的辅助因子与β'颌结构域相互作用，并且这种相互作用受到调控。
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