title: A microarray-based approach identifies ADP ribosylation factor-like protein 2 as a target of microRNA-16.
abstract: 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.
authors.affiliation: Jiangsu Diabetes Center, State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing, Jiangsu 210093, China.
authors.forename: Kehui
authors.initials: K
authors.lastname: Wang
authors.forename: Peng
authors.initials: P
authors.lastname: Li
authors.forename: Yanye
authors.initials: Y
authors.lastname: Dong
authors.forename: Xing
authors.initials: X
authors.lastname: Cai
authors.forename: Dongxia
authors.initials: D
authors.lastname: Hou
authors.forename: Jigang
authors.initials: J
authors.lastname: Guo
authors.forename: Yuan
authors.initials: Y
authors.lastname: Yin
authors.forename: Yujing
authors.initials: Y
authors.lastname: Zhang
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.forename: Hongwei
authors.initials: H
authors.lastname: Liang
authors.forename: Bowen
authors.initials: B
authors.lastname: Yu
authors.forename: Jiangning
authors.initials: J
authors.lastname: Chen
authors.forename: Ke
authors.initials: K
authors.lastname: Zen
authors.forename: Junfeng
authors.initials: J
authors.lastname: Zhang
authors.forename: Chen-Yu
authors.initials: CY
authors.lastname: Zhang
authors.forename: Xi
authors.initials: X
authors.lastname: Chen
mesh_terms: D020413:3' Untranslated Regions / Q000502:physiology*; D002453:Cell Cycle / Q000502:physiology*; D045744:Cell Line, Tumor; D019295:Computational Biology; D019204:GTP-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics; D020869:Gene Expression Profiling; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D006801:Humans; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis
chemical_list: D020413:3' Untranslated Regions; C505362:MIRN16 microRNA, human; D035683:MicroRNAs; C410130:ARL2 protein, human; D019204:GTP-Binding Proteins