In vitro analysis of the interaction between the small RNA SR1 and its primary target ahrC mRNA.
In vitro analysis of the interaction between the small RNA SR1 and its primary target ahrC mRNA.
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DOI：
10.1093/nar/gkm439
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发表时间：
2007
影响因子：
14.9
通讯作者：
Brantl S
中科院分区：
文献类型：
--
作者：
Heidrich N;Moll I;Brantl S
来自细菌染色体的小调控RNA（small regulatory RNAs，sRNAs）是近五年来研究的热点。然而，相对于研究充分的质粒编码的反义RNA，在结构要求、与其靶点相互作用的动力学和降解方面知之甚少。在这里，我们提出了一个详细的体外分析SR 1，枯草芽孢杆菌的sRNA，参与精氨酸催化剂的调节与其目标，ahrC mRNA的碱基配对。SR 1种类的不同长度和SR 1/ahrC RNA复合物的二级结构进行了测定，并缩小了复合物形成所需的功能片段。SR 1与其靶蛋白的初始接触包括SR 1终止子茎的5′端和ahrC转录起始位点下游100 bp的区域。对ahrC/SR 1复合物的足印研究和二级结构探测表明，SR 1通过诱导ahrC RBS下游的结构变化来抑制翻译起始。此外，证明了结合SR 1和ahrC RNA的Hfq不是促进ahrC/SR 1复合物形成所必需的，而是使ahrC mRNA能够翻译。计算了不同生长条件下SR 1的细胞内浓度。
Small regulatory RNAs (sRNAs) from bacterial chromosomes became the focus of research over the past five years. However, relatively little is known in terms of structural requirements, kinetics of interaction with their targets and degradation in contrast to well-studied plasmid-encoded antisense RNAs. Here, we present a detailed in vitro analysis of SR1, a sRNA of Bacillus subtilis that is involved in regulation of arginine catabolism by basepairing with its target, ahrC mRNA. The secondary structures of SR1 species of different lengths and of the SR1/ahrC RNA complex were determined and functional segments required for complex formation narrowed down. The initial contact between SR1 and its target was shown to involve the 5′ part of the SR1 terminator stem and a region 100 bp downstream from the ahrC transcriptional start site. Toeprinting studies and secondary structure probing of the ahrC/SR1 complex indicated that SR1 inhibits translation initiation by inducing structural changes downstream from the ahrC RBS. Furthermore, it was demonstrated that Hfq, which binds both SR1 and ahrC RNA was not required to promote ahrC/SR1 complex formation but to enable the translation of ahrC mRNA. The intracellular concentrations of SR1 were calculated under different growth conditions.
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影响因子：
14.9
作者：
Afonyushkin, T;Vecerek, B;Moll, I;Bläsi, U;Kaberdin, VR
通讯作者：
Kaberdin, VR
影响因子：
14.9
作者：
Mandin P;Repoila F;Vergassola M;Geissmann T;Cossart P
通讯作者：
Cossart P
影响因子：
3.6
作者：
Brantl, S;Wagner, EGH
通讯作者：
Wagner, EGH
DOI：
10.1073/pnas.170281497
发表时间：
2000-08-29
影响因子：
11.1
作者：
Lease, RA;Belfort, M
通讯作者：
Belfort, M
影响因子：
4.8
作者：
Nakamura, K;Yahagi, S;Yamane, K
通讯作者：
Yamane, K