SnoRNA-guided modifications of mRNA

SnoRNA 引导的 mRNA 修饰

基本信息

批准号：
10063945
负责人：
Christopher Lee Holley
金额：
$ 32.2万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-12-01 至 2023-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10063945
关键词：
Affinity Biochemical Biology Cell Line Cells Cellular biology Complement Complex Consumption Data Development Disease Enzymes Gene Expression Genetic Models Genetic Transcription Goals Guide RNA Half-Life Health Human Immunoprecipitation In Vitro Knock-out Label Lead Link Malignant Neoplasms Mass Spectrum Analysis Measures Mediating Messenger RNA Metabolic Methods Methylation Modeling Modification Mutation Analysis Organism Oxidative Stress Physiologic pulse Primer Extension Protein Biosynthesis Proteins RNA RNA Splicing RNA methylation Reactive Oxygen Species Reporting Ribosomal RNA Role Site Small Interfering RNA Small Nucleolar RNA Testing Thiouridine Time Translations Validation Work antisense nucleic acid base crosslink fibrillarin in vivo knock-down mRNA Expression overexpression peroxidasin protein expression recruit ribosome profiling stoichiometry transcriptome

项目摘要

ABSTRACT Recent studies have demonstrated that post-transcriptional mRNA modifications can regulate gene expression, with implications for cellular differentiation, development, and cancer. For example, the m6A modification can regulate fundamental aspects of mRNA biology, such as splicing, stability, and translation. In contrast, much less is known about internal 2'-O-methylation (Nm) on mRNA. Now, using genetic models, our data suggest that small nucleolar RNAs (snoRNAs) can guide internal Nm modification of mRNA, via the enzyme fibrillarin (FBL). Our findings also suggest that Nm modification can regulate mRNA levels and protein expression in vivo. In this application, we propose to define the mechanisms that lead to Nm modification of mRNA, and to understand how Nm regulates gene expression. Our long-term goals are to determine how snoRNA-guided modifications of mRNA impact gene expression, cell biology, and organism health. The objective of this proposal is to specifically determine how snoRNA-guided Nm modification regulates the expression of Pxdn mRNA. Our central hypothesis is that Rpl13a snoRNAs guide FBL-mediated Nm modification of Pxdn mRNA, and that the modification increases mRNA abundance but inhibits protein translation. We propose to test our hypothesis with the following Specific Aims: (1) Define the snoRNA(s) and protein components that target Pxdn mRNA for Nm modification; (2) Define how snoRNA-guided modification of Pxdn regulates RNA and protein expression; and (3) Discover the network of snoRNA-guided Nm modifications in cells. Successful completion of these Aims will define, for the first time, how Nm modification of mRNA regulates gene expression. We also expect that our work will define a new role for snoRNAs in the modification of mRNA. As such, this work will open new avenues in the study of both RNA modifications and snoRNA biology.

抽象的最近的研究表明，转录后 mRNA 修饰可以调节基因表达，对细胞分化、发育和癌症具有影响。例如，m6A 修饰可以调节 mRNA 生物学的基本方面，例如剪接、稳定性和翻译。相比之下就少了很多已知 mRNA 上的内部 2'-O-甲基化 (Nm)。现在，使用遗传模型，我们的数据表明，小核仁 RNA (snoRNA) 可以通过纤维蛋白 (FBL) 酶引导 mRNA 的内部 Nm 修饰。我们的研究结果还表明，Nm 修饰可以调节体内 mRNA 水平和蛋白质表达。在这个应用中，我们建议定义导致 mRNA Nm 修饰的机制，并了解 Nm 如何调节基因表达。我们的长期目标是确定 snoRNA 引导的修饰如何 mRNA 影响基因表达、细胞生物学和生物体健康。该提案的目的是具体确定 snoRNA 引导的 Nm 修饰如何调节 Pxdn mRNA 的表达。我们的中心假设 Rpl13a snoRNA 指导 FBL 介导的 Pxdn mRNA 的 Nm 修饰，并且该修饰增加 mRNA 丰度，但抑制蛋白质翻译。我们建议用以下方法来检验我们的假设具体目标： (1) 定义针对 Pxdn mRNA 进行 Nm 修饰的 snoRNA 和蛋白质成分； (2) 定义snoRNA引导的Pxdn修饰如何调节RNA和蛋白表达； (3) 发现细胞中 snoRNA 引导的 Nm 修饰网络。成功完成这些目标将定义，首次研究mRNA的Nm修饰如何调控基因表达。我们还希望我们的工作能够定义 snoRNA 在 mRNA 修饰中的新作用。因此，这项工作将为研究开辟新途径 RNA 修饰和 snoRNA 生物学。