Uncovering the regulatory principles of dynamic mRNA methylation

揭示动态mRNA甲基化的调控原理

• 批准号: 10675002
• 负责人: Sigrid H Nachtergaele
• 金额: $ 40.19万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10675002
• 关键词: Address; Area; Biochemical; Biochemistry; Biological Models; Biology; Cells; Chemicals; Complex; Data; Defect; Deposition; Development; Disease; Enzymes; Gene Expression Regulation; Genetic Transcription; Glean; High-Throughput Nucleotide Sequencing; Label; Laboratories; Malignant Neoplasms; Maps; Mediating; Messenger RNA; Methods; Methylation; Modification; Molecular; Neurodegenerative Disorders; Positioning Attribute; Postdoctoral Fellow; Prevalence; RNA; RNA Polymerase II; RNA Processing; RNA Splicing; RNA methylation; Regulation; Research; Site; System; Transcript; Translations; Work; human disease; innovation; molecular modeling; prevent; recruit; trafficking; transcriptome

Project Summary Chemical modification of RNA is a critical mechanism of gene expression regulation, controlling RNA processing, stability, and, in the case of mRNA, translation. The chemical diversity of RNA modifications suggests that there is extensive biology yet to be uncovered. A recent surge in this field has been driven largely by rapid advances in high throughput sequencing methods that allow us to map these marks on a transcriptome-wide scale. However, the majority of studies remain correlative and are not able to reveal the molecular mechanisms of RNA modification function or regulation. The information we glean from such studies often represents an average across all transcripts in the cell, and does not take into account the spatial organization or temporal control of RNA transcription, processing, and trafficking. Moreover, the majority of work on mRNA modifications has focused on a single modification, N6-methyladenosine (m6A), which represents only one of over one hundred modifications annotated to date. Here we describe two research directions in our laboratory that address fundamental questions about mRNA modifications. The first research direction aims to characterize a new mRNA modification, N1-methyladenosine (m1A). Since discovering this modification in mRNA in during my postdoctoral work, technical challenges have prevented us from understanding the functions and regulation of this mRNA modification. These challenges have also led to conflicting data and controversy over the presence and prevalence of m1A in mRNA. We have recently validated new m1A mRNA sites and we are now well positioned to finally shed light on this mysterious mRNA modification. The second research direction aims to identify the mechanisms that coordinate cotranscriptional modification of mRNAs as they are being synthesized. This remains mysterious for even the most well- characterized mRNA modification, m6A. We hypothesize that RNA polymerase II complexes recruit RNA modification enzymes during transcription, akin to how splicing and processing factors are cotranscriptionally recruited to nascent mRNA. Using detailed biochemistry and RNA labeling approaches, we will dissect the mechanisms of cotranscriptional RNA methylation. To tackle challenges such as these, our lab identifies specific settings, essentially molecular model systems, that allow us to dissect the molecular mechanisms of RNA modification-mediated regulation of gene expression. Once we reveal the regulatory components and principles that govern these specific systems, we then have a molecular foothold to determine how general or specific these principles are, and in which contexts they apply. Our work will open up a new areas of RNA biology and allow us to understand how defects in these mechanisms result in associated human diseases.

项目摘要 RNA的化学修饰是基因表达调控的关键机制， 加工，稳定性，以及mRNA的翻译。RNA修饰的化学多样性 表明还有大量的生物学有待发现。最近这一领域的激增主要是由 通过高通量测序方法的快速发展，使我们能够将这些标记定位在一个 转录组范围。然而，大多数研究仍然是相关的，不能揭示 RNA修饰功能或调节的分子机制。我们从这些研究中收集到的信息 通常代表细胞中所有转录物的平均值，并且不考虑空间分布。 RNA转录、加工和运输的组织或时间控制。而且大部分 mRNA修饰的工作集中在一种单一的修饰，N6-甲基腺苷（m6 A）， 仅代表迄今为止注释的一百多个修改之一。在这里，我们描述两个研究 在我们的实验室，解决有关mRNA修饰的基本问题的方向。第一次研究 方向旨在表征一种新的mRNA修饰，N1-甲基腺苷（m1A）。自从发现这个 在我的博士后工作中，技术挑战使我们无法 了解这种mRNA修饰的功能和调节。这些挑战也导致了 关于m1A在mRNA中的存在和流行的矛盾数据和争议。我们最近 验证了新的m1A mRNA位点，我们现在处于有利地位，最终揭示了这种神秘的mRNA 修改。第二个研究方向旨在确定协调共转录的机制 在mRNA合成时进行修饰。即使对最好的人来说，这仍然是神秘的- 特征性mRNA修饰，m6 A。我们假设RNA聚合酶II复合物募集RNA 转录过程中的修饰酶，类似于剪接和加工因子在转录过程中的作用。 招募到新生的mRNA中。使用详细的生物化学和RNA标记方法，我们将解剖 共同转录RNA甲基化的机制。为了应对这些挑战，我们的实验室 具体的设置，基本上是分子模型系统，使我们能够剖析的分子机制， RNA修饰介导的基因表达调控。一旦我们揭示了监管组件， 原则，支配这些具体的系统，然后我们有一个分子立足点，以确定如何一般或 这些原则是具体的，以及它们适用于什么情况。我们的工作将为RNA的研究开辟新的领域 生物学，并使我们了解这些机制的缺陷如何导致相关的人类疾病。