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酿酒酵母M1双链RNA病毒的SL1 RNA的溶液结构

Solution structure of the SL1 RNA of the M1 double-stranded RNA virus of Saccharomyces cerevisiae.

作者信息

Yoo J S, Cheong H K, Lee B J, Kim Y B, Cheong C

机构信息

Magnetic Resonance Team, Korea Basic Science Institute, Taejon 305-333, Korea.

出版信息

Biophys J. 2001 Apr;80(4):1957-66. doi: 10.1016/S0006-3495(01)76165-6.

Abstract

The 20-nucleotide SL1 VBS RNA, 5'-GGAGACGC[GAUUC]GCGCUCC (bulged A underlined and loop bases in brackets), plays a crucial role in viral particle binding to the plus strand and packaging of the RNA. Its structure was determined by NMR spectroscopy. Structure calculations gave a precisely defined structure, with an average pairwise root mean square deviation (RMSD) of 1.28 A for the entire molecule, 0.57 A for the loop region (C8-G14), and 0.46 A for the bulge region (G4-G7, C15-C17). Base stacking continues for three nucleotides on the 5' side of the loop. The final structure contains a single hydrogen bond involving the guanine imino proton and the carbonyl O(2) of the cytosine between the nucleotides on the 5' and 3' ends of the loop, although they do not form a Watson-Crick base pair. All three pyrimidine bases in the loop point toward the major groove, which implies that Cap-Pol protein may recognize the major groove of the SL1 loop region. The bulged A5 residue is stacked in the stem, but nuclear Overhauser enhancements (NOEs) suggest that A5 spends part of the time in the bulged-out conformation. The rigid conformation of the upper stem and loop regions may allow the SL1 VBS RNA to interact with Cap-Pol protein without drastically changing its own conformation.

摘要

20个核苷酸的SL1 VBS RNA,5'-GGAGACGC[GAUUC]GCGCUCC(下划线表示凸起的A,括号内为环区碱基),在病毒颗粒与正链的结合以及RNA的包装过程中发挥着关键作用。其结构通过核磁共振光谱法确定。结构计算得出了一个精确界定的结构,整个分子的平均成对均方根偏差(RMSD)为1.28 Å,环区(C8 - G14)为0.57 Å,凸起区(G4 - G7,C15 - C17)为0.46 Å。环区5'侧的三个核苷酸持续存在碱基堆积。最终结构包含一个氢键,涉及环区5'端和3'端核苷酸之间鸟嘌呤亚氨基质子与胞嘧啶的羰基O(2),尽管它们并未形成沃森-克里克碱基对。环区的所有三个嘧啶碱基都指向大沟,这意味着Cap-Pol蛋白可能识别SL1环区的大沟。凸起的A5残基堆积在茎中,但核Overhauser效应(NOE)表明A5部分时间处于凸起构象。上部茎区和环区的刚性构象可能使SL1 VBS RNA在不显著改变自身构象的情况下与Cap-Pol蛋白相互作用。

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