Structural features of the binding site for ribosomal protein S8 in Escherichia coli 16S rRNA defined using NMR spectroscopy.
Structural features of the binding site for ribosomal protein S8 in Escherichia coli 16S rRNA defined using NMR spectroscopy.
复制标题
使用 NMR 光谱确定大肠杆菌 16S rRNA 中核糖体蛋白 S8 结合位点的结构特征。
DOI：
10.1073/pnas.94.6.2139
复制
发表时间：
1997
影响因子：
11.1
通讯作者：
Nikonowicz,EP
中科院分区：
文献类型：
--
作者：
Kalurachchi,K;Uma,K;Zimmermann,RA;Nikonowicz,EP
大肠杆菌核糖体蛋白S8通过与16 SrRNA相互作用，在30 S核糖体亚基组装中起关键作用。S8还通过与spcoperon mRNA的相互作用参与核糖体蛋白表达的翻译调节。16 S rRNA内蛋白S8的结合位点包括核苷酸G588至G604和C634至C651，并且由两个碱基配对的螺旋区组成，所述螺旋区位于包含9个残基的遗传学保守核心元件的侧翼。我们研究了S8的rRNA结合位点的结构，无论是在游离状态下，并在蛋白质的存在下，使用NMR光谱。这两个螺旋片段的完整性得到了验证，并证实了从比较分析中预测的保守核心内存在G597·C643和A596·U644碱基对。此外，我们还鉴定了核心内由残基A595·（A596· U644）组成的碱基三联体。NMR数据表明，S8-RNA相互作用在RNA无显著变化的情况下完成。尽管如此，S8结合促进了U 598·A640碱基对的形成，并且似乎稳定了G597·C643和A596·U644碱基对。
Ribosomal protein S8 ofEscherichia coliplays a key role in 30S ribosomal subunit assembly through its interaction with 16S rRNA. S8 also participates in the translational regulation of ribosomal protein expression through its interaction withspcoperon mRNA. The binding site for protein S8 within the 16S rRNA encompasses nucleotides G588 to G604 and C634 to C651 and is composed of two base paired helical regions that flank a phylogenetically conserved core element containing nine residues. We have investigated the structure of the rRNA binding site for S8 both in the free state and in the presence of protein using NMR spectroscopy. The integrity of the two helical segments has been verified, and the presence of G597·C643 and A596·U644 base pairs within the conserved core, predicted from comparative analysis, have been confirmed. In addition, we have identified a base triple within the core that is composed of residues A595·(A596· U644). The NMR data suggest that S8–RNA interaction is accomplished without significant changes in the RNA. Nonetheless, S8 binding promotes formation of the U598·A640 base pair and appears to stabilize the G597·C643 and A596·U644 base pairs.