Identification and Characterization of Small Molecule-RNA Conjugates in Yeast

描述(申请人提供)：在过去的几十年里，广泛的功能RNA的发现揭示了RNA在诸如基因表达和蛋白质生物合成等基本细胞过程中的作用日益扩大；然而，RNA可用的已知化学多样性仍然相对有限。传统上，在生物系统中发现化学修饰的核糖核苷酸一直局限于基于靶点的方法；RNA分子的类别或核苷酸修饰是事先已知的，并在有限的背景下进行研究。最近，报道了一种发现小分子-RNA偶联物的一般筛查方法。与以前的方法不同，这种筛选不依赖于特定的小分子结构，不依赖于特定类别的RNA，也不依赖于结合物的特定生物功能。新的RNA化学修饰的发现有可能改善我们在基因调控或基因剪接等领域对RNA生物学的相对较差的理解，更具挑衅性的是，可能识别RNA在细胞中的全新角色，如新的催化、信号或基因调控模式。以前，这种方法的应用导致在两个细菌生物体中发现了两个新的5‘小分子-RNA偶联物。在这里，筛选将被用来鉴定酵母中存在的修饰核苷酸。在鉴定之后，重点将放在通过开发修饰的RNA的测序方法以及试图将修饰的RNA与已知的生物过程联系起来来表征修饰的核苷酸的生物学功能。这项拟议的研究有可能提供更全面的了解酵母中细胞RNA的化学多样性，并可能显著扩大我们对RNA的生物学甚至化学功能的理解。 与公共卫生相关：在过去的几十年里，细胞RNA的已知生物学作用已经大大扩大；然而，RNA的化学多样性相对保持不变。本研究旨在从酵母这一重要的真核模式生物中鉴定新的修饰RNA，并对其生物学功能进行表征。增加对RNA化学多样性的了解将潜在地提高对RNA生物学作用的理解，并最终导致新的治疗靶点。

DESCRIPTION (provided by applicant): Over the past few decades, the discovery of a wide range of functional RNAs has revealed an expanding role for RNA in essential cellular processes such as gene expression and protein biosynthesis; however, the known chemical diversity available to RNA is still relatively limited. Traditionally, the discovery of chemically modified ribonucleotides in biological systems has been limited to a target based approach; either the class of RNA molecules or the nucleotide modification were known beforehand and studied in a limited context. Recently, a general screen has been reported for the discovery of small molecule-RNA conjugates. Unlike previous methods, this screen is not dependent on a specific small-molecule structure, on a certain class of RNA, or on a particular biological function of the conjugate. The identification of novel chemical modifications of RNA has the potential to improve our still relatively poor understanding of RNA biology in areas such as gene regulation or gene splicing, and, more provocatively, may identify entirely new roles for RNA in the cell, such as new modes of catalysis, signaling, or gene regulation. Previously, application of this approach led to the discovery of two novel 5' small molecule-RNA conjugates in two bacterial organisms. Here, the screen will be used to identify modified nucleotides present in yeast. Following identification, focus will be placed on characterizing the biological function of the modified nucleotides through development of sequencing methods for modified RNAs as well as attempting to link the modified RNA to a known biological process. The proposed research has the potential to provide a more complete understanding of the chemical diversity of cellular RNA in yeast, and possibly significantly expand our understanding of the biological, and perhaps even chemical, functions of RNA. PUBLIC HEALTH RELEVANCE: The known biological roles of cellular RNA have expanded considerably in the past few decades; however, the chemical diversity of RNA has remained relatively constant. This proposal seeks to identify novel modified RNA in yeast, an important eukaryotic model organism, and characterize their biological function. An increased understanding of the chemical diversity of RNA will potentially improve understanding of the biological role of RNA, and, eventually, lead to new therapeutic targets.