GENETIC REGULATION THROUGH STRUCTURAL STUDIES OF RIBOSWITCH-METABOLITE COMPLEXES

通过核糖开关代谢物复合物的结构研究进行遗传调控

• 批准号: 8360016
• 负责人: JEFF SOUKUP
• 金额: $ 5.07万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8360016
• 关键词: Agonist; Antibiotic Resistance; Antibiotics; Bacteria; Bacterial Infections; Base Pairing; Binding; Binding Sites; Complex; Crystallization; Development; Distal; Elements; Funding; Gene Expression Regulation; Genes; Genetic; Grant; Guanine; Ligands; Metabolic; Metabolism; Molecular Probes; National Center for Research Resources; Nebraska; Principal Investigator; Process; RNA; Regulation; Research; Research Infrastructure; Resources; Source; United States National Institutes of Health; X-Ray Crystallography; analog; aptamer; base; combat; cost; design; fighting; functional genomics; novel; novel strategies

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The emergence of antibiotic resistance has required that new approaches be applied in order to effectively fight a host of medically relevant bacterial infections. The currently used, imprecise antibiotics need to be replaced with novel, rigorous, and safe treatments in order to combat the evolved bacterium of today. One way to destroy bacteria is to target one of their most essential processes, metabolism. The discovery of RNA structural elements, termed riboswitches, that bind cellular metabolites and control expression of essential metabolic genes provides a unique and distinct target for development of artificial agonists to fight bacterial infections. In order to rationally design and develop effective artificial agonists/antibiotics that target bacterial riboswitches, an understanding of the structural and functional details of the riboswitch-metabolite complex is essential. The aims of this grant focus on (1) probing the molecular contribution of a conserved base pair distal to the metabolite binding site within a riboswitch that binds guanine, (2) designing a crystallization construct for structural characterization of the newly discovered pre-queuosine1 riboswitch, and (3) determining the molecular interactions between the metabolite pre-queuosine1 and its riboswitch aptamer domain. The studies described here will provide atomic level detail of the interactions between riboswitches and their ligands. Structural studies of riboswitches are essential in order to gain detailed information about how the RNA interacts with its metabolite and to ultimately design non-natural metabolite analogs that can act as antibiotics. The X-ray crystallography studies described here will also have a high impact on understanding RNA-based gene regulation.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 抗生素耐药性的出现要求采用新的方法来有效对抗许多医学相关的细菌感染。 目前使用的不精确的抗生素需要用新颖、严格且安全的治疗方法取代，以对抗当今进化的细菌。 消灭细菌的一种方法是针对细菌最重要的过程之一——新陈代谢。 RNA 结构元件（称为核糖开关）的发现，可以结合细胞代谢物并控制必需代谢基因的表达，为开发抗细菌感染的人工激动剂提供了独特且独特的靶标。 为了合理设计和开发针对细菌核糖开关的有效人工激动剂/抗生素，了解核糖开关-代谢物复合物的结构和功能细节至关重要。 该资助的目的集中在 (1) 探索结合鸟嘌呤的核糖开关内代谢物结合位点远端保守碱基对的分子贡献，(2) 设计结晶构建体，用于新发现的 pre-queuosine1 核糖开关的结构表征，(3) 确定代谢物 pre-queuosine1 与其核糖开关适体结构域之间的分子相互作用。这里描述的研究将提供核糖开关及其配体之间相互作用的原子水平细节。为了获得有关 RNA 如何与其代谢物相互作用的详细信息并最终设计可充当抗生素的非天然代谢物类似物，核糖开关的结构研究至关重要。这里描述的 X 射线晶体学研究也将对理解基于 RNA 的基因调控产生重大影响。