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DsrA小RNA为适应多种靶标调控而具有的重要构象可塑性。

The important conformational plasticity of DsrA sRNA for adapting multiple target regulation.

作者信息

Wu Pengzhi, Liu Xiaodan, Yang Lingna, Sun Yitong, Gong Qingguo, Wu Jihui, Shi Yunyu

机构信息

Hefei National Laboratory for Physical Sciences at Microscale and School of Life Sciences, University of Science and Technology of China, Anhui 230027, China.

出版信息

Nucleic Acids Res. 2017 Sep 19;45(16):9625-9639. doi: 10.1093/nar/gkx570.

Abstract

In bacteria, small non-coding RNAs (sRNAs) could function in gene regulations under variable stress responses. DsrA is an ∼90-nucleotide Hfq-dependent sRNA found in Escherichia coli. It regulates the translation and degradation of multiple mRNAs, such as rpoS, hns, mreB and rbsD mRNAs. However, its functional structure and particularly how it regulates multiple mRNAs remain obscure. Using NMR, we investigated the solution structures of the full-length and isolated stem-loops of DsrA. We first solved the NMR structure of the first stem-loop (SL1), and further studied the melting process of the SL1 induced by the base-pairing with the rpoS mRNA and the A-form duplex formation of the DsrA/rpoS complex. The secondary structure of the second stem-loop (SL2) was also determined, which contains a lower stem and an upper stem with distinctive stability. Interestingly, two conformational states of SL2 in dynamic equilibrium were observed in our NMR spectra, suggesting that the conformational selection may occur during the base-pairing between DsrA and mRNAs. In summary, our study suggests that the conformational plasticity of DsrA may represent a special mechanism sRNA employed to deal with its multiple regulatory targets of mRNA.

摘要

在细菌中,小非编码RNA(sRNA)可在多种应激反应下发挥基因调控功能。DsrA是一种约90个核苷酸的、依赖于Hfq的sRNA,存在于大肠杆菌中。它调控多种mRNA的翻译和降解,如rpoS、hns、mreB和rbsD mRNA。然而,其功能结构,尤其是它如何调控多种mRNA仍不清楚。我们利用核磁共振(NMR)研究了DsrA全长及分离出的茎环结构。我们首先解析了第一个茎环(SL1)的NMR结构,并进一步研究了与rpoS mRNA碱基配对诱导的SL1解链过程以及DsrA/rpoS复合物的A-型双链体形成。还确定了第二个茎环(SL2)的二级结构,它包含一个稳定性不同的下部茎和上部茎。有趣的是,在我们的NMR谱中观察到SL2存在两种处于动态平衡的构象状态,这表明在DsrA与mRNA碱基配对过程中可能发生构象选择。总之,我们的研究表明,DsrA的构象可塑性可能代表了sRNA用于应对其多个mRNA调控靶点的一种特殊机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7a64/5766208/95b907acb17f/gkx570fig1.jpg

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