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（来自家长奖） 在细菌蛋白质合成过程中，30S 和 50S 核糖体亚基组装成 模板 mRNA 上具有翻译活性的 70S 核糖体。在革兰氏阳性人类细菌中 病原体金黄色葡萄球菌，一种称为冬眠-的单个小核糖体结合蛋白 促进因子 (SaHPF) 刺激 2.5-MDa 70S 单体二聚化，形成 平移沉默的 100S 复合体。 100S核糖体的生理功能仍然存在 这是一个谜，因为 100S 核糖体的时间丰度在不同个体之间差异很大 不同的细菌门，100S核糖体对翻译的整体影响是完全不同的 未知，不同细菌的 hpf 无效突变体缺乏共同的表型。而且，遥远的 相关的γ变形菌，例如大肠杆菌，需要两种蛋白质（EcRMF和EcHPF）来 实现100S复杂成型。我们小组的最新数据表明 SaHPF 是 对于老化金黄色葡萄球菌细胞中细菌的存活和核糖体库的维持至关重要。 令人惊讶的是，消除 Sa_hpf 只会导致翻译时基因子集的去抑制 引发。我们的目标是建立对 100S 功能的机械理解 核糖体的翻译能力和葡萄球菌发病机制。我们将采取多 学科方法涵盖遗传学、分子生物物理学、生物化学和整个动物 感染研究。目标 1 将确定可逆过程和涉及的因素 70S 和 100S 核糖体的转换。目标 2 将确定 SaHPF/100S 核糖体如何 以基因特异性方式抑制翻译。目标 3 将确定 100S 综合体在 核糖体周转和葡萄球菌病理生理学。这些目标有可能产生 对核糖体代谢的新见解，并激发持久性和慢性疾病的替代治疗方法 复发性葡萄球菌感染与长期生存密切相关 主机内部。