Project 6: Regulation and Function of Extended 3' UTR Transcripts in the Nervous System

项目6：扩展3UTR转录本在神经系统中的调控和功能

• 批准号: 9360973
• 负责人: Pedro Miura
• 金额: $ 21.6万
• 依托单位: UNIVERSITY OF NEVADA RENO
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9360973
• 关键词: 3' Untranslated Regions; Affect; Binding; Biogenesis; Biological; Biological Assay; Brain; Calcium Signaling; Calmodulin; Cells; Centers of Research Excellence; Chromatin; Chromatin Structure; Clustered Regularly Interspaced Short Palindromic Repeats; Communities; DNA Polymerase II; Data; Defect; Disease; Distal; Drosophila genus; ELAV protein; Enzymes; Event; Exhibits; Fluorescent in Situ Hybridization; Gene Chips; Generations; Genes; Genetic; Genetic Transcription; Heart Abnormalities; Hereditary Disease; Human; Human Genetics; Impairment; Length; Link; Maintenance; Messenger RNA; Methodology; MicroRNAs; Molecular; Morphology; Mus; Mutation; Nervous system structure; Neurophysiology - biologic function; Neurosciences; Organism; Pathway interactions; Pattern; Phenotype; Physiological; Play; Polyadenylation; Process; Protein Isoforms; RNA; RNA interference screen; RNA-Binding Proteins; Regulation; Reporter; Research; Role; Semaphorins; Signal Transduction; Site; Technology; Testing; Tissues; Transcript; Transcriptional Elongation Factors; Transgenes; Translations; Untranslated Regions; Work; axon guidance; chromatin modification; experimental study; fly; genome editing; human disease; in vivo; insight; mRNA Precursor; mutant; nervous system development; nervous system disorder; neurodevelopment; novel; relating to nervous system

PROJECT SUMMARY/ABSTRACT Regulation and Function of Extended 3´ UTR Transcripts in the Nervous System More than 50% of genes in diverse organisms undergo Alternative Cleavage and PolyAdenylation (APA) to generate multiple 3´ UTR mRNA isoforms. Thousands of novel extended 3´ UTRs have been recently identified to be preferentially expressed in the nervous system of fly, mouse and human. Such a pervasive and cell-specific event is likely to have wide-ranging physiologically relevant consequences for nervous system development, maintenance and disease. However, to date, few functional roles for extended 3´ UTRs have been identified. The long-term objectives are to uncover functions for these extended 3´ UTR isoforms in the nervous system, and elucidate the mechanisms of their biogenesis and biological activity. The focus is on a set of genes that play established roles in axon guidance. More than ten genes with roles in axon guidance express short and extended 3´ UTR isoforms, suggesting that APA is an important regulator for this key neurodevelopmental event. Some of these genes have direct relevance to human disease. For instance, mutations in calmodulin genes are implicated in multiple cardiac defects in humans. To investigate extended 3´ UTR function, CRISPR genome editing is employed to specifically delete these isoforms in Drosophila. This approach has been established, and preliminary work has uncovered that impairment of an extended 3´ UTR isoform, while leaving the short 3´ UTR isoforms intact, can impair nervous system development. In Aim 1, this approach is expanded to cam, the Drosophila calmodulin gene. In Aim 2, the role that chromatin modifications have on the biogenesis of extended 3´ UTRs is investigated. This builds upon ongoing work on the mechanism through which the ELAV regulates 3´ UTR extension. Overall, this work will establish APA as a crucial mechanism governing multiple genes that control axon guidance.

项目概要/摘要 神经系统中扩展 3´ UTR 转录本的调节和功能 不同生物体中超过 50% 的基因经过选择性切割和聚腺苷酸化 (APA) 生成多个 3´ UTR mRNA 亚型。最近有数千个新颖的扩展 3' UTR 确定优先在果蝇、小鼠和人类的神经系统中表达。如此普遍且 细胞特异性事件可能对神经系统产生广泛的生理相关后果 发育、维持和疾病。然而，迄今为止，扩展 3´ UTR 的功能角色还很少 已被识别。长期目标是揭示这些扩展的 3´ UTR 亚型在 神经系统，并阐明其生物发生和生物活性的机制。重点是一个集合 在轴突引导中发挥既定作用的基因。十多个在轴突引导中发挥作用的基因 表达短和延长的 3´ UTR 亚型，表明 APA 是该关键的重要调节因子 神经发育事件。其中一些基因与人类疾病直接相关。例如， 钙调蛋白基因突变与人类多种心脏缺陷有关。调查扩展 3´ UTR功能，CRISPR基因组编辑用于特异性删除果蝇中的这些亚型。这 方法已经建立，初步工作已发现扩展 3´ UTR 的损伤 亚型，在保持短 3' UTR 亚型完整的同时，可能会损害神经系统的发育。在目标 1 中，这 该方法扩展到果蝇钙调蛋白基因 cam。在目标 2 中，染色质修饰的作用 研究了延长 3´ UTR 的生物发生。这是建立在该机制正在进行的工作基础上的 ELAV 通过它调节 3´ UTR 延伸。总体而言，这项工作将使 APA 成为一个至关重要的 控制轴突引导的多个基因的机制。