Quantitative and mechanistic dissection of messenger RNA stability

RNA分子的稳定性可能有很大不同，这直接影响其功能，例如通过调节mRNAs产生的蛋白质数量。长RNA的稳定性是如何通过序列、化学修饰、结构和RNA结合蛋白(RBPs)的相互作用来确定的，以及这种稳定性如何根据RNA的来源而变化，目前尚不清楚。了解决定RNA稳定性的密码是理解基因表达调控如何在不同的生理和病理环境中重定向的关键，包括发育、传染病和肿瘤。这对于有效设计新的RNA疗法以及了解mRNAs和lncRNAs中的哪些突变可能产生有害后果也是重要的。在转录过程中安装的m6A RNA修饰最近已成为RNA稳定性的主要决定因素，由RNA结合蛋白(RBPs)和其他决定因素设置的额外决定因素仍不清楚，特别是对于不在细胞核中产生的RNA。在这个跨学科的联合提案中，我们的目标是从定量、功能、生物化学和结构上剖析序列元件、化学修饰、结构和限制性商业惯例如何影响mRNA的稳定性。它将最先进的高通量RNA分析和计算生物学(由Ulitsky和Schwartz实验室提供的专业知识)与尖端的生化和结构方法(由Meister和Fischer实验室提供)相结合，以揭示序列、加工、mRNA修饰和限制性商业惯例如何协同决定mRNA的稳定性。综上所述，这些研究将提供对决定mRNA生命周期的顺式和反式决定因素的详细机制理解。我们的研究也将对开发与生物医学及其他领域相关的新型RNA技术具有很高的价值。

RNA molecules can differ substantially in their stability, which directly impacts their function, e.g., through regulating the amount of protein produced by mRNAs. How the stability of long RNAs is determined via an interplay of sequence, chemical modifications, structure, and RNA binding proteins (RBPs), and how this varies depending on the origin of the RNA remains poorly understood. Understanding the codes determining RNA stability is key for understanding how regulation of gene expression is redirected in diverse physiological and pathological contexts, including development, infectious disease, and tumors. It is also important for effective design of new RNA therapies and for understanding which mutations in mRNAs and lncRNAs can have detrimental consequences. The m6A RNA modification, which is installed during transcription has recently emerged as a major determinant of RNA stability, with additional determinants set by RNA binding proteins (RBPs), andothers remaining unclear, in particular for RNAs that are are not produced in the nucleus. In this interdisciplinary joint proposal, we aim to quantitatively, functionally, biochemically, and structurally dissect how sequence elements, chemical modifications, structure and RBPs impact mRNA stability. It synergizes state-of-the art high-throughput RNA assays and computational biology (expertise provided by the Ulitsky and Schwartz labs), with cutting-edge biochemical and structural approaches (provided by the Meister and Fischer labs), to unravel how sequence, processing, mRNA modifications and RBPs synergize to dictate mRNA stability. Together, these studies will provide a detailed mechanistic understanding of the cis and trans determinants dictating the mRNA life cycle. Our investigations will also be highly valuable for developing novel RNA technologies relevant for biomedicine and beyond.