Alpha-synuclein mRNA is a putative microRNA target

小调控RNAs参与真核细胞的许多功能，microRNAs(MiRNAs)是动物中小调控RNAs的主要亚类，通过破坏和/或减少特定靶标mRNAs的翻译，在转录后调节基因表达。转录后基因调控具有革命性的意义，但人们对miRNAs在神经元或神经退行性疾病中所扮演的角色知之甚少。目的1：鉴定神经元样细胞中的多聚核糖体miRNP。与miRNA相关的生物化学知之甚少。我们假设含有miRNA的多聚核糖体核糖核蛋白复合体(MiRNPs)代表了miRNA：mRNA调节的生化底物。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，以表征这一重要颗粒的生化特性和蛋白质组成。目的2：鉴定和鉴定miRNA：mRNA对。虽然已知数百个人类miRNAs，但大多数mRNAs的靶标是未知的。我们发现了管理miRNA的“规则”：靶mRNA相互作用。利用这些指南，我们的生物信息学家合作者预测了受人类miRNAs调控的mRNA靶标。我们将使用细胞生物学工具来验证假设的与人类神经疾病相关的miRNA-mRNA合作伙伴。目的3：鉴定可能调节α-突触核蛋白(A-SN)表达的A miRNA相互作用。在初步证据的基础上，我们假设A-SN mRNA 3‘-非翻译区中进化保守的序列元件是由特定的miRNA(miR-93)识别的。A-SN蛋白在某些神经退行性疾病中起核心作用 MiRNA的调控在神经生物学、神经退行性疾病和RNA生物学中具有重要意义。我们将使用神经元细胞系来研究这种相互作用，作为miRNA：mRNA验证的原型。我们还将把分析扩展到健康和疾病中的人脑组织。

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.