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首页> 外文期刊>Nucleic Acids Research >Structural elements of the 3'-terminal coat protein binding site in alfalfa mosaic virus RNAs
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Structural elements of the 3'-terminal coat protein binding site in alfalfa mosaic virus RNAs

机译：苜蓿花叶病毒RNAs 3'-末端外壳蛋白结合位点的结构元件

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摘要

The 3'-terminal of the three genomic RNAs of alfalfa mosaic virus (AIMV) and ilarviruses contain a number of AUGC-motifs separated by hairpin structures. Binding of coat protein (CP) to such elements in the RNAs is required to initiate infection of these viruses. Determinants for CP binding in the 3'-terminal 39 nucleotides (nt) of AIMV RNA 3 were analyzed by band-shift assays. From the 5'- to 3'-end this 39 nt sequence contains AUGC-motif 3, stem-loop structure 2 (STLP2), AUGC-motif 2, stem-loop structure 1 (STLP1) and AUGC-motif 1. A mutational analysis showed that all three AUGC-motifs were involved in CP binding. Mutation of the A- and U-residues of motifs 1 or 3 had no effect on CP binding but similar mutations in motif 2 abolished CP binding. A mutational analysis of the stem of STLP1 and STLP2 confirmed the importance of these hairpins for CP binding. Randomization of the sequence of the stems and loops of STLP1 and STLP2 had no effect on CP binding as long as the secondary structure was maintained. This indicates that the two hairpins are not involved in sequence-specific interactions with CP. They may function in a secondary structure- specific interaction with CP and/or in the assembly of the AUGC-motifs in a configuration required for CP binding.
机译：苜蓿花叶病毒（AIMV）和伊拉病毒的三个基因组RNA的3'端含有许多由发夹结构分隔的AUGC基序。需要外壳蛋白（CP）与RNA中的此类元素结合才能启动这些病毒的感染。通过带移测定分析了AIMV RNA 3的3'-端39个核苷酸（nt）中CP结合的决定因素。从5'端到3'端，该39 nt序列包含AUGC-基序3，茎环结构2（STLP2），AUGC-基序2，茎环结构1（STLP1）和AUGC-基序1。分析表明，所有三个AUGC基序均参与CP结合。基序1或3的A和U残基的突变对CP结合没有影响，但是基序2中类似的突变消除了CP结合。对STLP1和STLP2茎的突变分析证实了这些发夹对CP结合的重要性。只要保持二级结构，STLP1和STLP2茎和环序列的随机化对CP结合都没有影响。这表明这两个发夹不参与与CP的序列特异性相互作用。它们可以在与CP进行的特定于二级结构的交互中和/或在CP绑定所需的配置中在AUGC-基序的组装中起作用。

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