Structural bases of the functions of RNA-protein machines - Project 5

RNA-蛋白质机器功能的结构基础 - 项目 5

基本信息

批准号：
7782579
负责人：
SCOTT A STROBEL
金额：
$ 28.54万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-04-01 至 2014-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7782579
关键词：
2&apos -deoxyguanosine 5&apos -phosphate Active Sites Amino Acids Antibiotics Bacteria Bacterial Infections Base Pairing Binding Biochemical Biocontrols Biological Assay Biological Process Catalysis Catalytic RNA Cells Chemical Structure Chemicals Collection Complex Cyclic GMP Data Defect Drug Delivery Systems Electrophoretic Mobility Shift Assay Elements Environment Enzymes Excision Exons Gene Expression Gene Structure Generations Genes Glycine Goals Guanosine Health Human Intervention Introns Ions Ligand Binding Ligands Ligation Measures Mediating Membrane Metals Microbial Biofilms Mutagenesis Nucleotides Organism Pathway interactions Point Mutation Process Proteins RNA RNA Binding RNA Biochemistry RNA Degradation RNA Folding RNA Splicing Reaction Second Messenger Systems Series Signaling Molecule Site Specificity Spliceosomes Structure Testing Transcript Translation Initiation Virulence X-Ray Crystallography aptamer base biological systems catalyst cell motility chemical reaction digital human disease mRNA Precursor quorum sensing response second messenger sensor small molecule transcription termination

项目摘要

RNAs that recognize small molecule ligands provide a widespread regulatory mechanism in biological systems. This study focuses on three RNAs that respond to small molecules, but in very different ways. Studies will be performed on the group I self-splicing RNA, the cooperative glycine riboswitch and a newly discovered riboswitch responsive to the second messenger cyclic diguanosine monophosphate (c-diGMP). 1. RNA splicing in response to the small molecule substrate guanosine. RNA splicing is the removal of an intron and the simultaneous ligation of its flanking exons in the generation of mature cellular RNA molecules. While most introns are removed by the spliceosome, some introns are able to catalyze their own removal from the primary transcript. The discovery of the group I class of introns provided the first indication that not all enzymes are proteins. Our goal is to structurally characterize each step in an RNA splicing pathway in order to understand how RNA functions as a catalyst. 2. Control of gene expression by cooperative binding of the amino acid glycine. Riboswitches are dynamic RNA folding domains that bind specific metabolites and control gene expression by modulation of transcription termination, translation initiation or RNA degradation. The glycine riboswitch comprises two similar aptamer domains that each bind glycine, but do so cooperatively. The result is a digital RNA sensor that is particularly sensitive to glycine concentration. Our goal is to understand the structural basis of glycine binding and cooperativity. 3. Analysis of a riboswitch responsive to a second messenger in the cell. c-dGMP was recently identified as a ligand for a broad collection of riboswitches, termed GEMM riboswitches. Our goal is to understand the structural and biochemical basis of c-diGMP binding by this new class of RNA molecule.

识别小分子配体的RNA在生物领域提供了广泛的调节机制系统。这项研究重点关注三种对小分子做出反应的 RNA，但反应方式却截然不同。研究将针对 I 组自剪接 RNA、协作甘氨酸核糖开关和新的发现了对第二信使环二鸟苷单磷酸（c-diGMP）有反应的核糖开关。 1. RNA 剪接响应小分子底物鸟苷。 RNA剪接是去除成熟细胞 RNA 生成中的内含子及其侧翼外显子的同时连接分子。虽然大多数内含子被剪接体去除，但一些内含子能够催化自己的内含子从主要转录本中删除。 I 类内含子的发现提供了第一个迹象并非所有酶都是蛋白质。我们的目标是从结构上表征 RNA 剪接的每个步骤途径，以了解 RNA 如何发挥催化剂的作用。 2.通过氨基酸甘氨酸的协同结合来控制基因表达。核糖开关是动态 RNA 折叠结构域结合特定代谢物并通过调节转录终止、翻译起始或 RNA 降解。甘氨酸核糖开关包含两个类似的适体结构域各自结合甘氨酸，但协同作用。结果是数字 RNA 传感器对甘氨酸浓度特别敏感。我们的目标是了解甘氨酸的结构基础约束力和协作性。 3. 分析对细胞中第二信使有反应的核糖开关。最近发现了 c-dGMP 作为一系列核糖开关的配体，称为 GEMM 核糖开关。我们的目标是了解此类新型 RNA 分子结合 c-diGMP 的结构和生化基础。