GENETIC REGULATION THROUGH STRUCTURAL STUDIES OF RIBOSWITCH-METABOLITE COMPLEXES

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH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 中心，不一定是研究者的机构。 抗生素耐药性的出现要求应用新的方法，以有效地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的，严格的和安全的治疗方法来取代，以对抗今天进化的细菌。 消灭细菌的一种方法是针对它们最重要的过程之一，新陈代谢。RNA结构元件（称为核糖开关）的发现为开发对抗细菌感染的人工激动剂提供了独特和独特的靶点，核糖开关结合细胞代谢物并控制必需代谢基因的表达。 为了合理地设计和开发有效的人工激动剂/抗生素的目标细菌核糖开关，核糖开关代谢物复合物的结构和功能的细节的理解是必不可少的。 该资助的目的集中在（1）探测结合鸟嘌呤的核糖开关内代谢物结合位点远端的保守碱基对的分子贡献，（2）设计用于新发现的前体核糖开关1的结构表征的结晶构建体，以及（3）确定代谢物前体核糖开关1及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和它们的配体之间的相互作用的原子水平的细节。核糖开关的结构研究是必不可少的，以获得有关RNA如何与其代谢物相互作用的详细信息，并最终设计出可以作为抗生素的非天然代谢物类似物。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。

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. 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.