Structure-based functional analysis of RNA Polymerase

RNA 聚合酶基于结构的功能分析

• 批准号: 2884826
• 金额: --
• 依托单位: Newcastle University
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2884826
• 关键词: Structure; based; functional; analysis; RNA

Research project summaryGene expression, the basic requirement for life, is considered the central dogma of molecular biology. This dogma describes the directional flow of genetic information from deoxyribonucleic acid (DNA) to ribonucleic acid (RNA) (transcription), to polypeptide chains (translation) that undergo post-translational modification to form functional proteins. Bacterial RNA polymerase (RNAp) is the central enzyme of gene expression whereby it accomplishes the polymerization of ribonucleoside triphosphates (NTPs). The catalytic core enzyme consists of five subunits In bacteria, RNAp, interacts with other cellular machineries, such as translation, DNA repair and replication. Bacterial RNAp is also a potent target for the development of antibacterials. RNAps in all living organisms are multisubunit, multidomain enzymes, and the functions of all the domains are still not well understood. Our laboratory solved structures of multisubunit RNAp from Escherichia coli (E.coli) stalled at a DNA lesion, which highlighted movements of some particular domains, which may be involved in signalling to translation and DNA repair machineries. Our laboratory also solved the structure of a small single-subunit relative of multi-subunit RNAps, which highlighted the structural conservation of domains of the active center, but divergence in the rest of the molecule, thus prompting the investigation of structure-based functional comparisons.Aims and objectivesThe overall aim of the project is to develop and characterize new inhibitors of RNAp. The project proposes to perform structure-based functional analysis of multisubunit RNAp and its single-subunit relative to bring insight into: the cross-talk between RNAp, ribosome, and DNA repair factors; functional conservation of the RNAp structure, principles of transcription inhibition by other cellular machineries and small molecules.The project will apply a wide arsenal of biochemical, microbiological, genetics, and structural biology techniques. We will use a range of known and new inhibitors of transcription as molecular probes of the functions of RNAps. Besides fundamental insights into the functions of bacterial RNAp, the project may lead to the discovery of new targets for inhibitors and novel candidate molecules for future development as antibacterials.KEYWORDSCell biology: understanding the fundamental properties, structure and function of the cell and how it responds to and influences its local environmentStructural biology and biophysics: understanding the atomic organisation of molecules and macromolecular complexes and the dynamic, functional relationships between these components in cells and biological systemsAntimicrobial resistance

基因表达是生命的基本要求，被认为是分子生物学的中心法则。这个教条描述了遗传信息从脱氧核糖核酸（DNA）到核糖核酸（RNA）（转录），再到多肽链（翻译）的定向流动，多肽链经过翻译后修饰形成功能蛋白质。细菌RNA聚合酶（RNAp）是基因表达的中心酶，通过它完成核糖核苷三磷酸（NTPs）的聚合。在细菌中，催化核心酶由五个亚基组成，RNA β与其他细胞机制相互作用，如翻译，DNA修复和复制。细菌RNAp也是开发抗菌药物的有效靶点。所有生物体中的RNAps都是多亚基、多结构域的酶，并且所有结构域的功能仍然没有很好地理解。我们的实验室解决了来自大肠杆菌（E.coli）的多亚基RNAp在DNA损伤处停滞的结构，这突出了一些特定结构域的运动，这些结构域可能参与翻译和DNA修复机制的信号传导。我们的实验室还解决了一个小的单亚基相对的多亚基RNAps的结构，这突出了活性中心的结构保守性，但在分子的其余部分的分歧，从而促进了基于结构的功能比较的调查。目的和objectivesThe项目的总体目标是开发和表征新的抑制剂的RNAp。本计画将应用生物化学、微生物学、遗传学、结构生物学等技术，对多亚基RNA p及其单亚基RNA p进行结构功能分析，以了解RNA p、核糖体、DNA修复因子之间的相互作用、RNA p结构的功能保守性、其他细胞机制和小分子抑制转录的原理等。我们将使用一系列已知的和新的转录抑制剂作为RNAps功能的分子探针。除了对细菌RNAp功能的基本认识外，该项目还可能导致发现抑制剂的新靶点和未来开发为抗菌剂的新型候选分子。关键词细胞生物学：了解细胞的基本性质，结构和功能以及它如何响应和影响其局部环境结构生物学和生物物理学：了解分子和大分子复合物的原子组织以及细胞和生物系统中这些组分之间的动态功能关系抗菌素耐药性