A non-canonical plant microRNA target site.
A non-canonical plant microRNA target site.
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DOI：
10.1093/nar/gku157
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发表时间：
2014-04
影响因子：
14.9
通讯作者：
Bouché N
中科院分区：
文献类型：
--
作者：
Brousse C;Liu Q;Beauclair L;Deremetz A;Axtell MJ;Bouché N
植物microRNAs （miRNAs）通常与它们的靶标形成近乎完美的双链，并介导mRNA的切割。在这里，我们描述了拟南芥中miR398的一个非常规miRNA靶标，一个编码蓝铜结合蛋白（BCBP）的mRNA。BCBP mRNA在其5 ‘ -非翻译区（UTR）携带一个miR398互补位点，与miRNA的5 ’区相反，有6个核苷酸凸起。尽管被认为对功能特别关键的靶位区域被破坏，但与传统的植物靶位一样，bbcp mrna在与miRNA相反的核苷酸10和11之间被ARGONAUTE1切割。在缺乏该miRNA的突变体或过表达该miRNA的转基因植物中，bbcbp mrna的水平与miR398的水平呈负相关。引入两个突变，破坏切割位点周围的miRNA互补性，使靶标具有抗切割性。BCBP位点在BCBP mRNA的环境之外发挥作用，不依赖于5 ' -UTR的位置。减少凸起不会干扰mir398介导的调节，完全去除凸起会提高切片效率。降解数据分析和目标预测显示，mir398 - bbp相互作用似乎是相当独特的。然而，我们的研究结果表明，在一个古老的保守miRNA的5 '区存在非完美配对的功能靶点。
Plant microRNAs (miRNAs) typically form near-perfect duplexes with their targets and mediate mRNA cleavage. Here, we describe an unconventional miRNA target of miR398 in Arabidopsis, an mRNA encoding the blue copper-binding protein (BCBP). BCBP mRNA carries an miR398 complementary site in its 5′-untranslated region (UTR) with a bulge of six nucleotides opposite to the 5′ region of the miRNA. Despite the disruption of a target site region thought to be especially critical for function, BCBP mRNAs are cleaved by ARGONAUTE1 between nucleotides 10th and 11th, opposite to the miRNA, like conventional plant target sites. Levels of BCBP mRNAs are inversely correlated to levels of miR398 in mutants lacking the miRNA, or transgenic plants overexpressing it. Introducing two mutations that disrupt the miRNA complementarity around the cleavage site renders the target cleavage-resistant. The BCBP site functions outside of the context of the BCBP mRNA and does not depend on 5′-UTR location. Reducing the bulge does not interfere with miR398-mediated regulation and completely removing it increases the efficiency of the slicing. Analysis of degradome data and target predictions revealed that the miR398-BCBP interaction seems to be rather unique. Nevertheless, our results imply that functional target sites with non-perfect pairings in the 5′ region of an ancient conserved miRNA exist in plants.
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通讯作者：
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