Opening and closing of the bacterial RNA polymerase clamp.

Opening and closing of the bacterial RNA polymerase clamp.
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DOI:
10.1126/science.1218716
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发表时间:
2012-08-03
期刊:
Science (New York, N.Y.)
影响因子:
--
通讯作者:
Ebright RH
Ebright RH
中科院分区:
其他
文献类型:
--
作者:
Chakraborty A;Wang D;Ebright YW;Korlann Y;Kortkhonjia E;Kim T;Chowdhury S;Wigneshweraraj S;Irschik H;Jansen R;Nixon BT;Knight J;Weiss S;Ebright RH

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使用单分子荧光共振能量转移,我们已经定义了细菌RNA聚合酶(RNAP)钳形成在转录起始和延伸的每一步。我们发现,钳主要是开放的自由RNAP和早期的中间体在转录起始,但关闭后,形成一个有催化能力的转录起始复合物,并保持关闭在初始转录和转录延伸。我们发现,四RNAP抑制剂干扰钳开放。我们建议,钳开放允许DNA加载到RNAP活性中心裂缝和解绕,DNA加载和解绕触发钳关闭,钳关闭占起始复合物的高稳定性和延伸复合物的高稳定性和持续合成能力。
Using single-molecule fluorescence resonance energy transfer, we have defined bacterial RNA polymerase (RNAP) clamp formation at each step in transcription initiation and elongation. We find that the clamp predominantly is open in free RNAP and early intermediates in transcription initiation, but closes upon formation of a catalytically competent transcription initiation complex and remains closed during initial transcription and transcription elongation. We show that four RNAP inhibitors interfere with clamp opening. We propose that clamp opening allows DNA to be loaded into and unwound in the RNAP active-center cleft, that DNA loading and unwinding trigger clamp closure, and that clamp closure accounts for the high stability of initiation complexes and the high stability and processivity of elongation complexes.
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