Administrative Equipment Supplement for Regulation and function of bacterial 100S ribosome

细菌 100S 核糖体调节和功能的管理设备补充剂

• 批准号: 10582284
• 负责人: Mee-Ngan F Yap
• 金额: $ 5.91万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2022-09-11
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10582284
• 关键词: Aging; Animals; Award; Bacteria; Bacterial Proteins; Binding Proteins; Biochemistry; Biophysics; Cells; Complex; Data; Dimerization; Distant; Equipment; Escherichia coli; Functional disorder; Gammaproteobacteria; Genes; Genetic; Genus staphylococcus; Goals; Hibernation; Human; Infection; Link; Maintenance; Messenger RNA; Metabolism; Molecular; Pathogenesis; Phenotype; Physiological; Probiotics; Process; Protein Biosynthesis; Proteins; Regulation; Relapse; Ribosomes; Role; Staphylococcal Infections; Staphylococcus aureus; Translations; derepression; insight; interdisciplinary approach; monomer; mutant; novel; pathogenic bacteria; preventive intervention

PROJECT SUMMARY (from the parental award) 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 (SaHPF) 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 (EcRMF and EcHPF) to achieve 100S complex formation. Recent data from our group demonstrate that SaHPF is essential for the bacterial survival and maintenance of the ribosome pool in aging S. aureus cells. Surprisingly, eliminating Sa_hpf 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 SaHPF/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.

项目摘要（摘自父母奖） 在细菌蛋白合成期间，30s和50s核糖体亚基组装到 模板mRNA上的翻译活性70S核糖体。在革兰氏阳性的人类细菌中 病原体金黄色葡萄球菌，一种单一的小核糖体结合蛋白，称为Hibernation- 促进因子（SAHPF）刺激2.5-mDa 70S单体的二聚化形成 翻译无声的100s综合体。 100S核糖体的生理功能仍然存在 由于100s核糖体的时间丰度在杂物中有很大差异 不同的细菌门，100s核糖体对翻译的全球影响完全是 未知的不同细菌的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（ECRMF和ECHPF） 实现100s复合物的形成。我们小组的最新数据表明SAHPF是 衰老的金黄色葡萄球菌细胞中核糖体池的细菌存活和维持所必需的。 出乎意料的是，消除SA_HPF导致仅在转化处的基因子集取决 引发。我们的目标是建立对100s功能的机械理解 核糖体的转化能力和葡萄球菌发病机理。我们将进行多个 跨越遗传学，分子生物物理学，生物化学和全动物的纪律方法 感染研究。 AIM 1将确定可逆的过程和因素 70s和100s核糖体的转化。 AIM 2将确定SAHPF/100S核糖体的方式 以基因特异性方式抑制翻译。 AIM 3将确定100s综合体的作用 核糖体更新和葡萄球菌病理生理学。这些目标有可能生产 对核糖体代谢的新颖见解，并激发了对持续和的替代治疗方法 复发的葡萄球菌感染与长期生存密切相关 主机内部。