Alpha-synuclein mRNA is a putative microRNA target

小调控rna参与许多真核细胞功能，动物物种中小调控rna的主要亚类microRNAs （miRNAs）通过破坏和/或减少特定“靶”mrna的翻译来调节转录后基因表达。转录后基因调控具有革命性意义，但mirna在神经元或神经退行性疾病中的作用知之甚少。目的1：表征神经元样细胞中的多核糖体miRNP。mirna相关的生物化学尚不清楚。我们假设含有miRNA的多核糖体核糖核蛋白复合物（miRNPs）代表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核糖体miRNP，以表征这一重要颗粒的生化特性和蛋白质成分。目的2：鉴定和表征miRNA:mRNA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的“规则”：靶mRNA相互作用。使用这些指南，我们的生物信息学家合作者预测由人类mirna调节的mRNA靶标。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：描述可能调节α -突触核蛋白（A- sn）表达的miRNA相互作用。在初步证据的基础上，我们假设a - sn mRNA的3'-非翻译区中一个进化上保守的序列元件被特定的miRNA （miR-93）识别。a - sn蛋白在一些神经退行性疾病及其相关疾病中起核心作用

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.