VISUALIZING ALLOSTERY IN THE GENE-REGULATORY LYSINE RIBOSWITCH

基因调节赖氨酸核糖开关中变构的可视化

• 批准号: 7954921
• 负责人: Nathan Baird
• 金额: $ 0.65万
• 依托单位: ILLINOIS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-01-01 至 2009-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7954921
• 关键词: Anti-Bacterial Agents; Bacteria; Binding; Biophysics; Complement; Computer Retrieval of Information on Scientific Projects Database; Drug Delivery Systems; Eukaryota; Event; Flavin Mononucleotide; Funding; Gene Expression; Glycine; Goals; Grant; Homeostasis; Institution; Lysine; Modeling; Molecular Conformation; RNA; Regulator Genes; Reporting; Research; Research Personnel; Resources; Role; Shapes; Solutions; Source; Structure; Thiamine Pyrophosphate; United States National Institutes of Health; X-Ray Crystallography; aptamer; insight; interest; response

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Riboswitches are recently described RNA domains that directly bind to cellular metabolites and control gene expression in cis in bacteria and eukaryotes. Due to their role in cellular homeostasis and their potential as antibacterial drug targets, structural studies of riboswitches are of immediate interest. Crystal structures of several types of riboswitches bound to their cognate metabolites have been reported. However, the mechanism of riboswitch function is not well understood due to the lack of structural information regarding the metabolite-free conformation. The goal of this proposal is to probe the solution structure of the metabolite-free conformation and the metabolite-bound conformation of four riboswitches by SAXS: the thiamine pyrophosphate (TPP), flavin mononucleotide (FMN), glycine (Gly), and lysine (Lys) riboswitches. Previous studies of the glycine riboswitch have demonstrated the utility of SAXS studies (Lipfert et al, JMB, 2007) to investigate the global size and shape of these RNA aptamers. The conformational change induced upon binding metabolite is expected to be on the order of 5 - 20 Angstroms and is well suited for experimentation at the APS. Performing SAXS studies on both the metabolite-free and metabolite-bound conformations will provide insight into the global structural changes involved in the gene-regulatory response of riboswitches. Obtaining Rg and P(r) plots for these conformations will allow us to model the RNA before and after the gene-regulatory switching event. These studies will complement concurrent structure determination of the metabolite bound conformation by X-ray crystallography, providing a detailed structural explanation of riboswitch function.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 核糖开关是最近描述的 RNA 结构域，可直接与细胞代谢物结合并控制细菌和真核生物中顺式基因表达。由于核糖开关在细胞稳态中的作用及其作为抗菌药物靶点的潜力，核糖开关的结构研究引起了人们的直接兴趣。已报道了几种与其同源代谢物结合的核糖开关的晶体结构。然而，由于缺乏有关无代谢物构象的结构信息，核糖开关功能的机制尚不清楚。 本提案的目标是通过 SAXS 探测四种核糖开关的无代谢物构象和代谢物结合构象的溶液结构：焦磷酸硫胺素 (TPP)、黄素单核苷酸 (FMN)、甘氨酸 (Gly) 和赖氨酸。赖氨酸）核糖开关。先前对甘氨酸核糖开关的研究已经证明了 SAXS 研究（Lipfert 等人，JMB，2007）在研究这些 RNA 适体的整体大小和形状方面的效用。结合代谢物时诱导的构象变化预计约为 5 - 20 埃，非常适合 APS 实验。对无代谢物和代谢物结合构象进行 SAXS 研究将有助于深入了解核糖开关基因调控反应中涉及的整体结构变化。获得这些构象的 Rg 和 P(r) 图将使我们能够在基因调控转换事件之前和之后对 RNA 进行建模。这些研究将补充 X 射线晶体学对代谢物结合构象的同步结构测定，提供核糖开关功能的详细结构解释。