Mechanistic Insights into m6A-Mediated Regulation of Brain Development

m6A 介导的大脑发育调节的机制见解

基本信息

批准号：
10295195
负责人：
Kathryn D Meyer
金额：
$ 39.48万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-11-10 至 2023-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10295195
关键词：
3&apos Untranslated Regions Adenosine Affinity Binding Binding Proteins Biochemical Brain Cell Culture Techniques Cell Differentiation process Cell Line Cell Proliferation Coupled Defect Deposition Development Disease Embryo Enzymes Event Fishes Future Gene Expression Gene Expression Profiling Gene Targeting Genetic Translation Goals Health Human In Vitro Lead Mediating Messenger RNA Methylation Methyltransferase Modeling Modification Multiprotein Complexes Mus Nervous System Physiology Neuroglia Neurons Nucleotides Pathway interactions Play Prevalence Process Property Protein Family Proteins RNA RNA Binding RNA Splicing RNA-Binding Proteins Reader Regulation Regulatory Pathway Research Research Personnel Resolution Role Site Specificity Terminator Codon Tissues Transcription Process Translations base cell type epitranscriptomics experimental study fly in vivo insight interdisciplinary approach mRNA Export mRNA Stability nerve stem cell nervous system disorder neurodevelopment neurogenesis novel protein complex recruit response stem cell differentiation stem cell proliferation transcriptome

项目摘要

ABSTRACT N6-methyladenosine, or m6A, is a highly abundant base modification which was recently shown to be present in thousands of cellular mRNAs. Many of the functional roles of this mark are carried out by m6A-binding proteins, or “readers”, which bind m6A residues and contribute to various aspects of mRNA regulation, including mRNA export, stability, and translation. Depletion of the methyltransferase enzymes that catalyze m6A formation has been shown to disrupt stem cell proliferation and differentiation and to lead to developmental defects. In the brain, where m6A is particularly abundant, m6A depletion has been shown to disrupt neurogenesis and cause severe neurodevelopmental abnormalities. However, the mechanisms through which m6A regulates gene expression to control neurodevelopmental processes are poorly understood. In addition, the proteins mediating m6A function in the developing brain have not been explored. Here, we will investigate the function of a previously unknown m6A binding protein during brain development. First, we will uncover the key features that determine m6A binding specificity. Second, we will utilize m6A-depleted neuronal cell lines coupled with transcriptome-wide RNA binding studies to uncover the cellular mRNAs that are targeted by this reader through m6A. Third, we will use a combination of global gene expression profiling and gene targeting approaches to determine how m6A:reader interactions contribute to gene expression changes and neurodevelopment. Collectively, these studies will characterize a novel m6A reader in the brain and will provide important mechanistic insight into how m6A regulates brain development.

抽象的 N6-甲基腺苷（M6A）是一种高度丰富的基础修饰，最近显示出存在于成千上万的细胞mRNA。该标记的许多功能作用是通过M6A结合蛋白来执行的，或结合M6A保留并有助于mRNA调节的各个方面的“读者”，包括mRNA 出口，稳定和翻译。催化M6A形成的甲基转移酶的耗竭我们被证明会破坏干细胞的增殖和分化，并导致发育缺陷。在大脑，其中M6a特别丰富，已显示M6A部署会破坏神经发生并导致严重的神经发育异常。但是，M6A调节基因的机制控制神经发育过程的表达知之甚少。另外，蛋白质介导尚未探索发育中的M6A功能。在这里，我们将研究以前的功能在大脑发育过程中，未知的M6a结合蛋白。首先，我们将发现确定的关键功能 M6A结合特异性。其次，我们将利用贫血的神经元细胞系与整个转录组的结合 RNA结合研究以发现该读取器通过M6A靶向的细胞mRNA。第三，我们会的结合全球基因表达分析和基因靶向方法来确定如何 M6A：读取器相互作用有助于基因表达变化和神经发育。总的来说，这些研究将表征大脑中新型的M6A阅读器，并将提供重要的机械洞察力，以了解如何 M6A调节大脑发育。