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NusA 和 NusG 调控转录相关的转录速率和泡囊结构的重塑。

Correlative Transcription Rate and Bubble Conformation Remodeled by NusA and NusG.

机构信息

MOE Key Laboratory of Laser Life Science and Institute of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou, Guangdong 510631, China.

Guangdong Key Laboratory of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou, Guangdong 510631, China.

出版信息

J Phys Chem B. 2023 Apr 6;127(13):2909-2917. doi: 10.1021/acs.jpcb.2c08771. Epub 2023 Mar 28.

Abstract

Transcription is highly regulated by a variety of transcription factors, among which NusA and NusG act contradictorily in (E. coli) that NusA stabilizes a paused RNA polymerase (RNAP) and NusG suppresses it. The mechanism of the NusA and NusG regulations on RNAP transcription has been addressed, but their effect on the conformational changes of the transcription bubble correlated with transcription kinetics remains elusive. By using single-molecule magnetic trap, we identify a reduction in the transcription rate of ∼40% events by NusA. Although the rest ∼60% of transcription events exhibit unaffected transcription rates, a NusA-enhanced standard deviation of the transcription rate is observed. NusA remodeling also increases the extent of DNA unwinding in the transcription bubble by 1-2 base pairs, which can be reduced by NusG. The NusG remodeling is more significant on the RNAP molecules with reduced transcription rates rather than those without. Our results provide a quantitative view on the mechanisms of transcriptional regulation by NusA and NusG factors.

摘要

转录受到多种转录因子的高度调控,其中 NusA 和 NusG 在大肠杆菌中表现出矛盾的作用,NusA 稳定暂停的 RNA 聚合酶(RNAP),而 NusG 则抑制它。NusA 和 NusG 对 RNAP 转录的调控机制已经得到解决,但它们对与转录动力学相关的转录泡构象变化的影响仍不清楚。通过使用单分子磁阱,我们发现 NusA 使转录率降低了约 40%。尽管其余约 60%的转录事件表现出未受影响的转录率,但观察到 NusA 增强了转录率的标准偏差。NusA 重塑还增加了转录泡中 DNA 解链的程度 1-2 个碱基对,NusG 可以减少这种解链。NusG 重塑在转录率降低的 RNAP 分子上更为显著,而不是在没有降低转录率的分子上更为显著。我们的结果提供了关于 NusA 和 NusG 因子转录调控机制的定量观点。

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