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首页> 外文期刊>Biochemistry >Ensemble analysis of primary microRNA structure reveals an extensive capacity to deform near the drosha cleavage site
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Ensemble analysis of primary microRNA structure reveals an extensive capacity to deform near the drosha cleavage site

机译：初级microRNA结构的集合分析显示，在drosha裂解位点附近具有广泛的变形能力

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摘要

Most noncoding RNAs function properly only when folded into complex three-dimensional (3D) structures, but the experimental determination of these structures remains challenging. Understanding of primary microRNA (miRNA) maturation is currently limited by a lack of determined structures for nonprocessed forms of the RNA. SHAPE chemistry efficiently determines RNA secondary structural information with single-nucleotide resolution, providing constraints suitable for input into MC-Pipeline for refinement of 3D structure models. Here we combine these approaches to analyze three structurally diverse primary microRNAs, revealing deviations from canonical double-stranded RNA structure in the stem adjacent to the Drosha cut site for all three. The necessity of these deformable sites for efficient processing is demonstrated through Drosha processing assays. The structure models generated herein support the hypothesis that deformable sequences spaced roughly once per turn of A-form helix, created by noncanonical structure elements, combine with the necessary single-stranded RNA-double-stranded RNA junction to define the correct Drosha cleavage site.
机译：大多数非编码RNA仅在折叠成复杂的三维（3D）结构后才能正常运行，但是对这些结构的实验确定仍具有挑战性。目前，由于缺乏针对非加工形式的RNA的确定结构，对初级microRNA（miRNA）成熟的了解受到限制。 SHAPE化学可有效确定具有单核苷酸分辨率的RNA二级结构信息，从而提供适合输入MC-Pipeline进行优化3D结构模型的约束条件。在这里，我们结合这些方法来分析三种结构上不同的一级microRNA，揭示出这三个在与Drosha切割位点相邻的茎中均偏离规范的双链RNA结构。通过Drosha加工试验证明了这些可变形部位有效加工的必要性。本文生成的结构模型支持以下假设：由非规范结构元素产生的每转A型螺旋大约每圈间隔一次的可变形序列与必要的单链RNA-双链RNA连接相结合，以定义正确的Drosha切割位点。

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