Widespread regulatory activity of vertebrate microRNA* species
Widespread regulatory activity of vertebrate microRNA* species
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DOI：
10.1261/rna.2537911
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发表时间：
2011-02-01
期刊：
影响因子：
4.5
通讯作者：
Lai, Eric C.
中科院分区：
文献类型：
--
作者：
Yang, Jr-Shiuan;Phillips, Michael D.;Lai, Eric C.
microRNA（miRNA）生物发生过程中的专性中间体是一种类似于22个核苷酸的RNA双链体，成熟的miRNA优先掺入沉默复合物中。其伴侣miRNA* 种类通常被认为是乘客RNA，其调节能力尚未在脊椎动物中进行系统研究。我们的生物信息学分析表明，相当一部分的miRNA* 物种在脊椎动物进化过程中是严格保守的，在它们的种子区域中共同表现出最大的保守性，并定义了互补基序，其在脊椎动物3 '-UTR进化过程中的保守性具有统计学意义。对22个miRNA表达构建体的功能测试显示，大多数可以抑制miRNA和miRNA* 完美匹配报告基因，并且miRNA：miRNA* 传感器抑制的比率与miRNA：miRNA* 读数的内源性比率相关。微阵列数据的分析提供了转录组范围的证据，表明与肿瘤发生相关的几种miRNAs，包括mir-17、mir-34 a和mir-19的成熟和星星链种类的种子匹配靶标的调节。最后，3 '-UTR传感器测定和诱变测试证实了通过星星种子位点直接抑制5个miR-19* 靶标。总体而言，我们的数据表明，miRNA* 物种对脊椎动物调控网络有明显的影响，在研究miRNA功能及其对疾病状态的贡献时应考虑到这一点。
An obligate intermediate during microRNA (miRNA) biogenesis is an similar to 22-nucleotide RNA duplex, from which the mature miRNA is preferentially incorporated into a silencing complex. Its partner miRNA* species is generally regarded as a passenger RNA, whose regulatory capacity has not been systematically examined in vertebrates. Our bioinformatic analyses demonstrate that a substantial fraction of miRNA* species are stringently conserved over vertebrate evolution, collectively exhibit greatest conservation in their seed regions, and define complementary motifs whose conservation across vertebrate 3'-UTR evolution is statistically significant. Functional tests of 22 miRNA expression constructs revealed that a majority could repress both miRNA and miRNA* perfect match reporters, and the ratio of miRNA: miRNA* sensor repression was correlated with the endogenous ratio of miRNA: miRNA* reads. Analysis of microarray data provided transcriptome-wide evidence for the regulation of seed-matched targets for both mature and star strand species of several miRNAs relevant to oncogenesis, including mir-17, mir-34a, and mir-19. Finally, 3'-UTR sensor assays and mutagenesis tests confirmed direct repression of five miR-19* targets via star seed sites. Overall, our data demonstrate that miRNA* species have demonstrable impact on vertebrate regulatory networks and should be taken into account in studies of miRNA functions and their contribution to disease states.