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首页> 外文期刊>Journal of Molecular Biology >Interaction of the Bacillus stearothermophilus ribosomal protein S15 with its 5 '-translational operator mRNA
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Interaction of the Bacillus stearothermophilus ribosomal protein S15 with its 5 '-translational operator mRNA

机译：嗜热脂肪芽孢杆菌核糖体蛋白S15与5'-翻译操纵子mRNA的相互作用

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摘要

The Bacillus stearothermophilus ribosomal protein S15 (BS15) binds both a three-helix junction in the central domain of 16 S ribosomal RNA and its cognate mRNA. Native gel mobility-shift assays show that BS15 interacts specifically and with high affinity to the F-untranslated region (5'-UTR) of tl-tis cognate mRNA with an apparent dissociation constant of 3(+/-0.3) nM. In order to localize the structural elements that are essential for BS15 recognition, a series of deletion mutants of the full cognate mRNA were prepared and tested in the same gel-shift assay. The minimal binding site for BS15 is a 50 nucleotide RNA showing a close secondary structure resemblance to the BS15 binding region from 16 S rRNA. There are two major structural motifs that must be maintained for high-affinity binding. The first being a purine-rich three-helix junction, and the second being an internal loop. The sequence identity of the internal loops differs greatly between the BS15 mRNA and rRNA sites, and this difference is correlated to discrimination between wild-type BS15 and a BS15(H45R) mutant. The association and dissociation kinetics measured for the 5'-UTR-BS15 interaction are quite slow, but are typical for a ribosomal protein-RNA interaction. The BS15 mRNA and 16 S rRNA binding sites share a common secondary structure yet have little sequence identity. The mRNA and rRNA may in fact present similar if not identical structural elements that confer BS15 recognition.
机译：嗜热脂肪芽孢杆菌核糖体蛋白S15（BS15）结合16 S核糖体RNA中央结构域中的三螺旋连接点及其同源mRNA。天然凝胶迁移率位移分析表明BS15特异性相互作用，并与tl-tis同源mRNA的F非翻译区（5'-UTR）具有高亲和力，表观解离常数为3（+/- 0.3）nM。为了定位对于BS15识别至关重要的结构元件，制备了一系列完整同源mRNA的缺失突变体，并在相同的凝胶位移试验中进行了测试。 BS15的最小结合位点是一个50核苷酸的RNA，显示出与16 S rRNA的BS15结合区非常相似的二级结构。高亲和力结合必须保留两个主要的结构基序。第一个是富含嘌呤的三螺旋连接，第二个是内部回路。内部环的序列同一性在BS15 mRNA和rRNA位点之间存在很大差异，并且这种差异与野生型BS15和BS15（H45R）突变体之间的区分有关。测量的5'-UTR-BS15相互作用的缔合和解离动力学相当慢，但对于核糖体蛋白-RNA相互作用而言是典型的。 BS15 mRNA和16 S rRNA结合位点具有相同的二级结构，但几乎没有序列同一性。实际上，即使不相同，mRNA和rRNA也可能呈现相似的结构，从而赋予BS15识别能力。

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