Regulation and function of bacterial 100S ribosome

细菌100S核糖体的调控和功能

基本信息

批准号：
9930945
负责人：
Mee-Ngan F Yap
金额：
$ 33.18万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-08-01 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9930945
关键词：
Address Aging Animals Anti-Bacterial Agents Bacteria Bacterial Proteins Binding Binding Proteins Biochemical Biochemistry Biogenesis Biology Biophysics Cells Chronic Complex Computer Simulation Cyanobacterium Data Dimerization Dissociation Distant Equilibrium Escherichia coli Firmicutes Fluorescence Polarization Functional disorder Gammaproteobacteria Genes Genetic Genetic Transcription Genus staphylococcus Goals Growth Guanosine Guanosine Triphosphate Hibernation Human Infection Kinetics Knock-out Label Lead Link Maintenance Mediating Messenger RNA Metabolism Molecular Mutagenesis Pathogenesis Phase Phenotype Physiologic pulse Physiological Preventive Intervention Probiotics Process Production Protein Biosynthesis Proteins Regulation Regulatory Pathway Relapse Resistance Ribonucleases Ribosomes Role Sigma Factor Staphylococcal Infections Staphylococcus aureus Staphylococcus aureus infection Structure System Testing Translation Initiation Translations Up-Regulation Uridine Variant analog attenuation crosslink derepression dimer exhaust genome-wide host colonization in vivo insight interdisciplinary approach light scattering logarithm monomer mouse model mutant novel pathogen pathogenic bacteria prevent ribosome profiling

项目摘要

SUMMARY During bacterial protein synthesis, the 30S and 50S ribosomal subunits assemble into the translationally active 70S ribosome on template mRNA. In the Gram-positive human bacterial pathogen Staphylococcus aureus, a single small ribosome-binding protein called hibernation-promoting factor (HPFSa) stimulates the dimerization of 2.5-MDa 70S monomers to form the translationally silent 100S complex. The physiological function of the 100S ribosome remains enigmatic because the temporal abundance of the 100S ribosome varies considerably among different bacterial phyla, the global impact of the 100S ribosome on translation is completely unknown, and hpf null mutants of different bacteria lack a common phenotype. Moreover, distantly related gammaproteobacteria, such as E. coli, require two proteins (RMFEc and HPFEc) to achieve 100S complex formation. Recent data from our group demonstrate that HPFSa is essential for the bacterial survival and maintenance of the ribosome pool in aging S. aureus cells. Surprisingly, eliminating hpfSa causes the derepression of only a subset of genes at translational initiation. Our goal is to establish a mechanistic understanding of the function of the 100S ribosome in translational capacity and staphylococcal pathogenesis. We will take a multi-disciplinary approach that spans genetics, molecular biophysics, biochemistry, and whole animal infection studies. Aim 1 will determine the process and factors involved in the reversible conversion of 70S and 100S ribosomes. Aim 2 will determine how the HPFSa/100S ribosome inhibits translation in a gene- specific manner. Aim 3 will identify the roles of the 100S complex in ribosome turnover and staphylococcal pathophysiology. These aims have the potential to produce novel insights into ribosome metabolism and inspire alternate treatments for persistent and relapsed staphylococcal infections that are intimately linked to survive for an extended period inside the host.

概括在细菌蛋白合成过程中，30s和50s核糖体亚基组装成翻译模板mRNA上的活跃的70S核糖体。在革兰氏阳性的人类细菌病原体葡萄球菌中金黄色葡萄球菌，一种称为冬眠促进因子（HPFSA）的单个小核糖体结合蛋白刺激 2.5-mDa 70S单体的二聚化形成翻译静音的100s复合物。生理 100s核糖体的功能仍然神秘，因为100s核糖体的时间丰度在不同细菌门中有很大不同，100s核糖体对翻译的全球影响是完全未知，不同细菌的HPF无效突变体缺乏常见的表型。而且，遥不可及相关的γ-杆菌，例如大肠杆菌，需要两种蛋白质（RMFEC和HPFEC）才能达到100s 复合形成。我们小组的最新数据表明，HPFSA对于细菌生存至关重要以及在衰老的金黄色葡萄球菌细胞中维持核糖体池。令人惊讶的是，消除HPFSA会导致在翻译起始中仅消除基因的一部分。我们的目标是建立机械了解100S核糖体在翻译能力和葡萄球菌发病机理中的功能。我们将采用跨越遗传学，分子生物物理学，生物化学和整个遗传学的多学科方法动物感染研究。 AIM 1将确定可逆转化的过程和因素 70秒和100s核糖体。 AIM 2将确定HPFSA/100S核糖体如何抑制基因中的翻译具体方式。 AIM 3将确定100S复合物在核糖体周转和葡萄球菌中的作用病理生理学。这些目标有可能对核糖体代谢和激发与持续和复发的葡萄球菌感染的替代治疗在主机内部长期生存。