Regulation of Messenger RNA Turnover in Mammalian Cells

哺乳动物细胞中信使 RNA 周转的调节

• 批准号: 10368955
• 负责人: Ann-Bin Shyu
• 金额: $ 45.75万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10368955
• 关键词: 3' Untranslated Regions; Allergic inflammation; Area; Autoimmune Diseases; Biology; Complex; Cytoplasm; Development; Event; Excision; Gene Expression; Individual; Kinetics; Laboratories; Link; Longevity; Malignant Neoplasms; Mammalian Cell; Messenger RNA; Metabolic; Modification; Pathologic; Pathway interactions; Phase; Phosphorylation; Play; Poly(A) Tail; Polyadenylation; Process; Protein Biosynthesis; Protein Isoforms; Proteins; RNA; RNA Degradation; Regulation; Research; Role; Shapes; Signal Transduction; Stimulus; Therapeutic Agents; human disease; mRNA Decay; mRNA Stability; novel therapeutics; protein complex; transcriptome

Project Description Research in my laboratory focuses on elucidating the principles and regulatory mechanisms that govern messenger RNA turnover in mammalian cells. mRNA turnover plays an essential role in regulating gene expression via control of mRNA stability and quality, both globally and at the level of individual mRNAs. In mammalian cells, all major modes of mRNA decay are triggered by deadenylation (i.e., the removal of the poly(A) tails from the 3' end of mRNAs), a rate-limiting process involving two consecutive kinetic phases. Although previous studies provided a clear picture of the mechanistic steps and participating factors of mRNA decay pathways, relatively little is known about the impact of deadenylation and its modulation on mRNA turnover at the transcriptome level. Particularly lacking is an understanding of how coordinated changes in stability for whole groups of mRNAs contribute to programming or reprogramming of the transcriptome when mammalian cells respond to intra- or extra-cellular stimuli. In the cytoplasm, mRNAs fulfill their functions in the form of mRNA-protein complexes (mRNPs). mRNPs are highly dynamic entities, being continuously remodeled by rapid exchanges of their protein constituents, dictating individual mRNAs' fates at each step of their lifespan. Any inappropriate remodeling of an mRNP complex has the potential to disrupt its proper engagement in downstream events. Currently, one exciting and underexplored area of research in RNA biology is the remodeling of mRNPs at individual, group, and global levels during the process of mRNA deadenylation and decay. Our present proposal focuses on three seemingly disparate aspects of mammalian RNA biology that are linked by their potential to shape the mammalian transcriptome through modulating global mRNA turnover and mRNP remodeling. These are: 1) Alternative 3' end processing and polyadenylation (APA), which generates mRNA isoforms with distinct 3' untranslated regions; 2) mRNA N6-methyladenosine (m6A) modification, which creates mRNA isoforms with different metabolic fate; and 3) Phosphorylation of ancillary deadenylation factors, which alters mRNA deadenylation and decay. In the past few years, we have made several key findings regarding these three areas that have helped lay the groundwork for the proposed studies in this application. We have also adapted and further developed key analytical approaches to elucidating the impacts of the targeted processes on mRNA turnover across the transcriptome and the mechanisms by which deadenylation impacts mRNA stability through mRNP remodeling. Successful completion of the proposed studies will offer a new framework for elucidating the signal-dependent regulation of mRNA stability and mRNP remodeling at the transcriptome level, and the results will significantly expand understanding of the fundamental principles governing eukaryotic mRNA turnover.

项目描述 我实验室的研究重点是阐明 控制哺乳动物细胞中信使RNA的周转。mRNA周转在调节 通过控制mRNA的稳定性和质量，在整体和个体水平上进行基因表达 mRNA。在哺乳动物细胞中，mRNA衰变的所有主要模式都是由去腺苷化（即，的 从mRNA的3'末端去除poly（A）尾），这是一个限速过程，涉及两个连续的 动力学阶段虽然以前的研究提供了一个清晰的图片的机械步骤， mRNA降解途径的参与因素，相对较少的是了解的影响，去腺苷化 及其在转录组水平上对mRNA周转的调节。尤其缺乏的是对 整组mRNA稳定性的协调变化如何有助于编程， 当哺乳动物细胞响应细胞内或细胞外刺激时，转录组的重编程。在 在细胞质中，mRNA以mRNA-蛋白质复合物（mRNP）的形式实现其功能。mRNP是 高度动态的实体，通过其蛋白质成分的快速交换而不断重塑， 决定着每个mRNA生命周期中的命运。任何mRNP的不适当重塑 复合体有可能破坏其在下游事件中的适当参与。目前，一个令人兴奋的 在RNA生物学中，尚未探索的研究领域是在个体、群体和群体中重塑mRNP。 在mRNA去腺苷化和衰变过程中的全局水平。 我们目前的建议集中在哺乳动物RNA生物学的三个看似不同的方面， 通过调节整体mRNA来塑造哺乳动物的转录组， 转换和mRNP重塑。这些是：1）替代的3'末端加工和聚腺苷酸化（阿帕）， 其产生具有不同3'非翻译区的mRNA同种型; 2）mRNA N6-甲基腺苷（m6 A） 修饰，其产生具有不同代谢命运的mRNA同种型;和3）磷酸化 辅助去腺苷化因子，其改变mRNA去腺苷化和衰变。在过去的几年里，我们 在这三个领域取得了几项重要发现，为 本申请中的建议研究。我们还调整并进一步开发了关键的分析 阐明靶向过程对mRNA周转的影响的方法 转录组和去腺苷化通过mRNP影响mRNA稳定性的机制 重塑成功完成拟议的研究将提供一个新的框架， 在转录组水平上对mRNA稳定性和mRNP重塑的信号依赖性调节， 这些结果将大大扩展对真核mRNA基本原理的理解， 周转