Alpha-synuclein mRNA is a putative microRNA target

小调控RNA参与许多真核细胞功能，microRNA（miRNAs）是动物物种中小调控RNA的主要亚类，其通过使特异性“靶”mRNA不稳定和/或减少其翻译来转录后调节基因表达。转录后基因调控具有革命性的意义，但对miRNAs在神经元或神经退行性疾病中所起的作用知之甚少。目的1：鉴定神经元样细胞中的多核糖体miRNP。miRNA相关的生物化学知之甚少。我们假设，含有miRNA的多核糖体核糖核蛋白复合物（miRNP）代表了miRNA：mRNA调控的生化底物。我们将从Weri视网膜母细胞瘤细胞中部分纯化多聚核糖体miRNP，以表征这个重要颗粒的生化特性和蛋白组分。AiM 2：鉴定和表征miRNA：mRNA配对。尽管已知数百种人类miRNA，但大多数mRNA靶点是未知的。我们发现了控制miRNA：靶mRNA相互作用的“规则”。使用这些指导方针，我们的生物信息学家合作者预测由人类miRNA调控的mRNA靶点。我们将使用细胞生物学工具来验证与人类神经系统疾病相关的假设miRNA-mRNA伴侣。目的3：研究一种可调控α-突触核蛋白（A-SN）表达的miRNA相互作用。基于初步证据，我们假设A-SN mRNA的3 '非翻译区中的进化保守序列元件被特定的miRNA（miR-93）识别。A-SN蛋白在一些神经退行性疾病中起着重要作用， 通过miRNA的调节将在神经生物学、神经变性疾病和RNA生物学中具有重要意义。我们将使用神经元细胞系研究这种相互作用，作为miRNA：mRNA验证的原型。我们还将分析扩展到健康和疾病的人脑组织。

Small regulatory RNAs participate in many eukaryotic cell functions, microRNAs (miRNAs), the major subclass of small regulatory RNAs in animal species, regulate gene expression post-transcriptionally by destabilizing and/or reducing the translation of specific 'target' mRNAs. Post-transcriptional gene regulation has revolutionary implications, yet very little is known of the roles played by miRNAs in neurons or neurodegenerative disease. AIM 1: CHARACTERIZE THE POLYRIBOSOMAL miRNP IN NEURON-LIKE CELLS. miRNA-related biochemistry is poorly understood. We hypothesize that miRNA-containing polyribosomal ribonucleoprotein complexes (miRNPs) represent the biochemical substrate for miRNA:mRNA regulation. We will partially purify the polyribosomal miRNP from Weri retinoblastoma cells, to characterize the biochemical properties and the protein components of this important particle. AiM 2: IDENTIFY AND CHARACTERIZE miRNA:mRNA PAIRS. Although hundreds of human miRNAs are known, most mRNA targets are unknown. We discerned "rules" that govern miRNA:target mRNA interaction. Using these guidelines our bioinformatician collaborators predict mRNA targets regulated by human miRNAs. We will use cell biological tools to verify hypothesized miRNA-mRNA partners relevant to human neurological diseases. AIM 3: CHARACTERIZE A miRNA INTERACTION THAT MAY REGULATE THE EXPRESSION OF ALPHA-SYNUCLEIN (A-SN). On the basis of preliminary evidence, we hypothesize that an evolutionarily conserved sequence element in the 3'-untranslated region of A-SN mRNA is recognized by a specific miRNA (miR-93). A-SN protein plays a central role in some neurodegenerative diseases and its regulation by a miRNA would have important implications in neurobiology, neurodegenerative disease, and RNA biology. We will study this interaction using neuronal cell lines, as a prototype of miRNA:mRNA validation. We will also extend the analyses to human brain tissue in health and disease.