Morosyuk S V, Cunningham P R, SantaLucia J
Department of Chemistry, Wayne State University, Detroit, MI 48202, USA.
J Mol Biol. 2001 Mar 16;307(1):197-211. doi: 10.1006/jmbi.2000.4431.
The solution structure of the conserved 690 hairpin from Escherichia coli 16 S rRNA was determined by NMR spectroscopy. The 690 loop is located at the surface of the 30 S subunit in the platform region and has been implicated in interactions with P-site bound tRNA, E-site mRNA, S11 binding, IF3 binding, and in RNA-RNA interactions with the 790 loop of 16 S rRNA and domain IV of 23 S rRNA. The structure reveals a novel sheared type G690.U697 base-pair with a single hydrogen bond from the G690 amino to U697-04. G691 and A696 also form a sheared pair and U692 forms a U-turn with an H-bond to the A695 non-bridging phosphate oxygen. The sheared pairs and U-turn result in the continuous single-stranded stacking of five residues from 6693 to U697 with their Watson-Crick functional groups exposed in the minor groove. The overall fold of the 690 hairpin is similar to the anticodon loop of tRNA. The structure provides an explanation for chemical protection patterns in the loop upon interaction with tRNA, the 50 S subunit, and S11. In vivo genetic studies demonstrate the functional importance of the motifs observed in the solution structure of the 690 hairpin.
通过核磁共振光谱法确定了来自大肠杆菌16S rRNA的保守690发夹结构。690环位于30S亚基平台区域的表面,参与与P位点结合的tRNA、E位点mRNA、S11结合、IF3结合以及与16S rRNA的790环和23S rRNA的结构域IV的RNA-RNA相互作用。该结构揭示了一种新型的剪切型G690.U697碱基对,从G690氨基到U697-04有一个氢键。G691和A696也形成一个剪切对,U692形成一个U型转弯,与A695非桥连磷酸氧形成氢键。剪切对和U型转弯导致从693到U697的五个残基连续单链堆积,其沃森-克里克官能团暴露在小沟中。690发夹的整体折叠与tRNA的反密码子环相似。该结构解释了与tRNA、50S亚基和S11相互作用时环中的化学保护模式。体内遗传学研究证明了在690发夹溶液结构中观察到的基序的功能重要性。