Ludwig Katelyn R, Dahl Richard, Hummon Amanda B
Harper Cancer Research Institute, University of Notre Dame , Notre Dame, Indiana 46617 United States.
Department of Microbiology and Immunology, Indiana University School of Medicine , South Bend, Indiana 46202 United States.
J Proteome Res. 2016 May 6;15(5):1497-505. doi: 10.1021/acs.jproteome.5b01101. Epub 2016 Apr 6.
MicroRNAs (miRNAs) are post-transcriptional regulators of gene expression that are implicated in a number of disease states. MiRNAs can exist as individual entities or may be clustered and transcribed as a single polycistron. The mirn23a cluster consists of three miRNAs: miR-23a, miR-24-2, and miR-27a. Although these miRNAs are transcribed together, they often exist at varying levels in the cell. Despite the fact that the mirn23a cluster is known to play a role in a number of diseases and developmental processes, few direct targets have been identified. In this study, we examined the effects of miR-23a, miR-24-2, miR-27a, or the mirn23a cluster overexpression on the proteome of 70Z/3 pre-B lymphoblast cells. Quantitative mass spectrometry using isobaric tags for relative and absolute quantification (iTRAQ) allowed for the global profiling of cell lines after miRNA overexpression. We identified a number of targets of each miRNA that contained predicted miRNA seed sequences and are likely direct targets. In addition, we discovered a cohort of shared miRNA targets and cluster targets, demonstrating the importance of studying miRNA clusters in their entirety.
微小RNA（miRNA）是基因表达的转录后调节因子，与多种疾病状态有关。miRNA可以作为单个实体存在，也可以成簇存在并作为单个多顺反子转录。mirn23a簇由三个miRNA组成：miR-23a、miR-24-2和miR-27a。尽管这些miRNA一起转录，但它们在细胞中的水平通常各不相同。尽管已知mirn23a簇在多种疾病和发育过程中起作用，但已确定的直接靶标很少。在本研究中，我们检测了miR-23a、miR-24-2、miR-27a或mirn23a簇过表达对70Z/3前B淋巴母细胞蛋白质组的影响。使用等压标签进行相对和绝对定量（iTRAQ）的定量质谱分析能够在miRNA过表达后对细胞系进行全面分析。我们鉴定出了每个miRNA的许多靶标，这些靶标包含预测的miRNA种子序列，可能是直接靶标。此外，我们还发现了一组共同的miRNA靶标和簇靶标，证明了整体研究miRNA簇的重要性。
