mRNA Methylation: a Novel Regulatory Mechanism in the Neuronal Transcriptome

描述(由申请者提供)：该项目旨在提高凯特·迈耶博士在过渡到独立研究科学家职业生涯时的技能、知识和科学培训。迈耶博士最近发表了一项里程碑式的研究，她首次证明了N6-甲基腺苷(M6A)是mRNA中广泛存在的可逆碱基修饰。M6A在mRNAs中表现出独特的分布，最高丰度的区域在终止密码子附近和5‘非编码区。此外，迈耶博士发现，M6A在大脑中特别丰富，其水平稳步上升 贯穿整个神经发育过程。这表明，信使核糖核酸甲基化是神经元信使核糖核酸调节的一种广泛的机制，有助于脑的发育和功能。然而，尽管m6A在大脑中普遍存在，并有可能影响神经元转录组的很大一部分，但其功能仍不清楚。本文提出的研究将利用技术和概念创新相结合的方法来研究m6A在神经元mRNAs中的定位、调节和功能。在目标1中，迈耶博士将开发一种新的工具，在单核苷酸分辨率下全球检测m6A残基，并将识别在大脑发育过程中动态甲基化的神经元mRNAs。目的2将利用生化实验和m6A定位技术的组合来确定控制神经元mRNAs中m6A定位的因素。这些实验将有助于回答长期存在的问题，即腺苷甲基化是如何定向到转录本的特定区域的。最后，目标3将探讨m6A在调节神经元mRNA翻译中的作用。这些实验的结果将为m6A如何促进基本大脑功能提供重要的见解，并将推动我们对有助于心理健康和疾病的分子和表观遗传学事件的理解。为了实现这些研究目标，迈耶博士将接受计算机编程和生物信息分析方面的广泛培训。这套新技能对迈耶博士来说将是无价的，因为作为分子神经生物学领域的独立研究人员，她的许多研究将涉及下一代大型测序数据集的生成和分析。除了让迈耶博士发展计算分析方面的专业知识外，这一奖项还将进一步提高她在拨款撰写、指导和其他关键领域的技能，这些领域对于独立研究科学家的成功职业生涯至关重要。因此，在该奖项的指导阶段结束时，迈耶博士将处于一个理想的位置，以实现她的长期职业目标，即经营一个成功的研究实验室，研究信使核糖核酸甲基化及其在心理健康和疾病中的作用。

DESCRIPTION (provided by applicant): This project is designed to enhance the skills, knowledge, and scientific training of Dr. Kate Meyer as she transitions into a career as an independent research scientist. Dr. Meyer recently published a landmark study in which she showed for the first time that N6-methyladenosine (m6A) is a widespread, reversible base modification in mRNA. m6A exhibits a unique distribution within mRNAs, with the most highly enriched regions being near the stop codon and in the 5'UTR. Additionally, Dr. Meyer discovered that m6A is particularly abundant within the brain and that its levels increase steadily throughout neurodevelopment. This suggests that mRNA methylation is a widespread mechanism of neuronal mRNA regulation which contributes to brain development and function. However, despite the prevalence of m6A within the brain and its potential to influence a substantial portion of the neuronal transcriptome, its function remains unknown. The research proposed here will utilize a combination of technically and conceptually innovative approaches to investigate the localization, regulation, and function of m6A in neuronal mRNAs. In Aim 1, Dr. Meyer will develop a new tool for globally detecting m6A residues at single-nucleotide resolution and will identify the neuronal mRNAs which are dynamically methylated during brain development. Aim 2 will utilize a combination of biochemical experiments and m6A mapping techniques to determine the factors that control m6A localization in neuronal mRNAs. These experiments will help answer the long-standing question of how adenosine methylation is directed to particular regions of a transcript. Finally, Aim 3 will explore the role of m6A in regulating neuronal mRNA translation. The results of these experiments will provide important insight into how m6A contributes to basic brain function and will propel our understanding of the molecular and epigenetic events that contribute to mental health and disease. To achieve these research goals, Dr. Meyer will receive extensive training in computer programming and bioinformatic analysis. This new skill set will be invaluable for Dr. Meyer to have, since many of her studies as an independent researcher in the field of molecular neurobiology will involve the generation and analysis of large next- generation sequencing datasets. In addition to enabling Dr. Meyer to develop expertise in computational analysis, this award will further enhance her skills in grant writing, mentoring, and other key areas which are crucial for a successful career as an independent research scientist. Thus, by the end of the mentored phase of this award, Dr. Meyer will be in an ideal position to achieve her long-term career goal of running a successful research laboratory studying mRNA methylation and its role in mental health and disease.