A Microarray-based Approach Identifies ADP Ribosylation Factor-like Protein 2 as a Target of microRNA-16
A Microarray-based Approach Identifies ADP Ribosylation Factor-like Protein 2 as a Target of microRNA-16
复制标题
基于微阵列的方法将 ADP 核糖基化因子样蛋白 2 识别为 microRNA-16 的靶标
DOI：
10.1074/jbc.m110.178335
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发表时间：
2011-03-18
影响因子：
4.8
通讯作者：
Chen, Xi
中科院分区：
文献类型：
--
作者：
Wang, Kehui;Li, Peng;Chen, Xi
microRNA（miRNAs）通常被认为通过mRNA降解或翻译抑制来负调节其靶基因的表达。在这里，我们展示了一种有效的方法来识别miRNA靶基因，通过筛选miRNA水平变化后的整体mRNA水平的变化。在这项研究中，我们使用mRNA微阵列来测量miR-16水平升高或降低的三种细胞系中的总体mRNA表达，并基于多靶点预测算法进行生物信息学分析。为了进一步研究预测的miR-16靶基因，我们选择了与miR-16表达模式相反的基因。进一步研究了可能与miR-16相互作用的候选靶基因之一ADP-核糖基化因子样蛋白2（ARL 2）。首先，通过计算预测，推断ARL 2是理想的miR-16靶标。其次，用miR-16前体处理显著消除了ARL 2 mRNA和蛋白水平，而miR-16抑制剂增加了ARL 2 mRNA和蛋白水平。第三，荧光素酶报告基因测定证实miR-16直接识别ARL 2的3 '-非翻译区（3'-UTR）。最后，我们发现miR-16可以调节增殖并诱导显著的G 0/G1细胞周期停滞，这至少部分是由于ARL 2的下调。总之，本研究表明，整合全球mRNA谱分析和生物信息学工具可能为进一步研究给定miRNA的潜在靶点提供基础。这些结果还说明了靶向ARL 2的miR-16在调节增殖和细胞周期进程中的新功能。
microRNAs (miRNAs) are generally thought to negatively regulate the expression of their target genes by mRNA degradation or by translation repression. Here we show an efficient way to identify miRNA target genes by screening alterations in global mRNA levels following changes in miRNA levels. In this study, we used mRNA microarrays to measure global mRNA expression in three cell lines with increased or decreased levels of miR-16 and performed bioinformatics analysis based on multiple target prediction algorithms. For further investigation among the predicted miR-16 target genes, we selected genes that show an expression pattern opposite to that of miR-16. One of the candidate target genes that may interact with miR-16, ADP-ribosylation factor-like protein 2 (ARL2), was further investigated. First, ARL2 was deduced to be an ideal miR-16 target by computational predictions. Second, ARL2 mRNA and protein levels were significantly abolished by treatment with miR-16 precursors, whereas a miR-16 inhibitor increased ARL2 mRNA and protein levels. Third, a luciferase reporter assay confirmed that miR-16 directly recognizes the 3'-untranslated region (3'-UTR) of ARL2. Finally, we showed that miR-16 could regulate proliferation and induce a significant G0/G1 cell cycle arrest, which was due at least in part, to the down-regulation of ARL2. In summary, the present study suggests that integrating global mRNA profiling and bioinformatics tools may provide the basis for further investigation of the potential targets of a given miRNA. These results also illustrate a novel function of miR-16 targeting ARL2 in modulating proliferation and cell cycle progression.