Pausing controls branching between productive and non-productive pathways during initial transcription in bacteria.
Pausing controls branching between productive and non-productive pathways during initial transcription in bacteria.
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DOI：
10.1038/s41467-018-03902-9
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发表时间：
2018-04-16
影响因子：
16.6
通讯作者：
Kapanidis AN
中科院分区：
文献类型：
--
作者：
Dulin D;Bauer DLV;Malinen AM;Bakermans JJW;Kaller M;Morichaud Z;Petushkov I;Depken M;Brodolin K;Kulbachinskiy A;Kapanidis AN
细菌中的转录受到多种分子机制的控制，这些分子机制精确调节基因表达。最近的研究表明，细菌 RNA 聚合酶 (RNAP) 的初始 RNA 合成会因暂停而中断；然而，暂停决定因素以及暂停与有效和失败 RNA 合成的关系仍然知之甚少。使用单分子 FRET 和生化分析，我们表明在 6-nt RNA (ITC6) 合成后 RNAP 遇到的暂停使得启动子逃逸强烈依赖于 NTP 浓度。从机制上讲，暂停的 ITC6 充当检查点，将 RNAP 引导至三种竞争途径之一：有效转录、失败的 RNA 释放或新的 unscrunching/scrunching 途径。启动子的循环收缩/收缩产生了一种长期存在的、RNA 结合的暂停状态；因此，失败的 RNA 释放和 DNA 整理并不像之前想象的那样紧密相连。最后，我们的新模型将暂停与初始转录的失败和生产结果结合起来。细菌 RNA 聚合酶的 RNA 合成会因暂停而中断，但人们对它们在 RNA 合成中的作用知之甚少。在这里，作者使用单分子 FRET 和生化分析来表明暂停调节初始转录的失败和生产结果之间的分支。
Transcription in bacteria is controlled by multiple molecular mechanisms that precisely regulate gene expression. It has been recently shown that initial RNA synthesis by the bacterial RNA polymerase (RNAP) is interrupted by pauses; however, the pausing determinants and the relationship of pausing with productive and abortive RNA synthesis remain poorly understood. Using single-molecule FRET and biochemical analysis, here we show that the pause encountered by RNAP after the synthesis of a 6-nt RNA (ITC6) renders the promoter escape strongly dependent on the NTP concentration. Mechanistically, the paused ITC6 acts as a checkpoint that directs RNAP to one of three competing pathways: productive transcription, abortive RNA release, or a new unscrunching/scrunching pathway. The cyclic unscrunching/scrunching of the promoter generates a long-lived, RNA-bound paused state; the abortive RNA release and DNA unscrunching are thus not as tightly linked as previously thought. Finally, our new model couples the pausing with the abortive and productive outcomes of initial transcription. RNA synthesis by bacterial RNA polymerase is interrupted by pauses but their role in RNA synthesis is poorly understood. Here the authors use single-molecule FRET and biochemical analysis to show that pausing regulates branching between the abortive and productive outcomes of initial transcription.
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影响因子：
12.3
作者：
Imashimizu M;Takahashi H;Oshima T;McIntosh C;Bubunenko M;Court DL;Kashlev M
通讯作者：
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作者：
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