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Regulation and function of bacterial hibernating 100S ribosome

细菌冬眠100S核糖体的调控和功能

基本信息

批准号：
10703477
负责人：
Mee-Ngan F Yap
金额：
$ 32.19万
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 2026-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10703477
关键词：
Address Area Bacteria Binding Biochemistry Biogenesis Biological Catalysis Cell Survival Cells Comparative Study Complex Cytoplasmic Protein Data Development Dimerization Endoribonucleases Escherichia coli Eukaryota Exonuclease Exoribonucleases Firmicutes Genetic Screening Goals Gram-Negative Bacteria Gram-Positive Bacteria Growth Guanosine Triphosphate Heterogeneity Hibernation Homologous Gene Hydrolysis Infection Intervention Knock-out Knowledge Life Link Location Longevity Mass Spectrum Analysis Messenger RNA Metabolic Microscopy Modeling Molecular Mutagenesis Organism Pathogenicity Pathway interactions Peptide Elongation Factor G Phase Phenotype Phosphodiesterase I Positioning Attribute Post-Translational Protein Processing Probiotics Process Proteins Quality Control RNA Regulation Resolution Resources Ribonucleases Ribosomal Proteins Ribosomal RNA Ribosomes Role Site Staphylococcus aureus Structure Time Transcription Process Translating Translation Initiation Translation Process Translational Derepression Translations Virulence Virulence Factors Work antimicrobial bacterial genetics cell growth chronic infection combat endonuclease genome-wide human pathogen improved in vivo innovation insight interdisciplinary approach microscopic imaging mutant novel pathogenic bacteria preservation prevent preventive intervention public health relevance rapid growth ribonuclease R segregation small molecule inhibitor spatiotemporal structural biology superresolution microscopy translation factor unpublished works

项目摘要

PROJECT SUMMARY Ribosome hibernation is a conserved mechanism used by both bacteria and eukaryotes to prevent translation and to extend organismal lifespan. Recent studies from various bacterial species, including pathogenic Staphylococcus aureus, have provided compelling evidence for a critical role of hibernating 100S ribosomes in protecting the ribosomal pool from damage, in addition to blocking translational initiation. We found that S. aureus ribosomes lacking hibernation-promoting factor (HPF) are rapidly degraded by the 3’-5’ exonuclease RNase R and other hitherto unknown ribonucleases. In our unpublished work, we isolated an additional ribonuclease mutant that rescues the loss of ribosomes in S. aureus. Surprisingly, we found that ribosomes are not the only target of S. aureus HPF; instead, HPF could interact with a cytoplasmic protein of unknown biological activity, thereby reducing the abundance of hibernating 100S ribosomes. We further demonstrated that HPF is restricted to a specific subcellular localization during rapid growth, providing a rare glimpse of possible HPF segregation from actively translating ribosomes. In this proposal, we will undertake a highly multidisciplinary approach consisting of structural biology, omics, bacterial genetics, biochemistry and high-resolution microscopy to achieve the following goals: (1) Determine the molecular mechanisms by which HPF protects ribosomes from ribonucleolytic cleavage. (2) Determine the previously undiscovered extraribosomal role of HPF. (3) Determine how the spatiotemporal localization of HPF avoids translation conflicts. HPF and RNase R are evolutionarily conserved virulence factors among nosocomial gram-positive and gram-negative bacteria, completion of these aims will provide significant mechanistic insight into innovative counterstrategies to combat recalcitrant infections by perturbing the biogenesis and turnover of hibernating ribosomes.

项目摘要核糖体冬眠是细菌和真核生物用来阻止翻译的一种保守机制并延长生物体的寿命。最近对各种细菌物种的研究，包括致病性金黄色葡萄球菌，提供了令人信服的证据，证明冬眠的100 S核糖体在除了阻断翻译起始外，还保护核糖体库免受损伤。我们发现S.金黄色缺乏冬眠促进因子（HPF）的核糖体被3 '-5'核酸外切酶RNase迅速降解 R和其他迄今未知的核糖核酸酶。在我们未发表的工作中，我们分离出一种额外的核糖核酸酶，突变体，挽救了S中核糖体的损失。金黄色。令人惊讶的是，我们发现核糖体不是唯一的 S的目标。金黄色葡萄球菌HPF;相反，HPF可以与未知生物活性的细胞质蛋白相互作用，从而减少冬眠的100 S核糖体的丰度。我们进一步证明了HPF是受限制的，在快速生长过程中的一个特定的亚细胞定位，提供了一个罕见的可能HPF分离一瞥核糖体的主动翻译。在这一建议中，我们将采取高度多学科的方法，包括结构生物学、组学、细菌遗传学、生物化学和高分辨率显微镜，实现以下目标：（1）确定HPF保护核糖体免受核糖核裂解(2)确定以前未发现的HPF的核糖体外作用。(3)确定 HPF的时空定位如何避免翻译冲突。HPF和RNase R在进化上保守的毒力因子之间的医院革兰氏阳性和革兰氏阴性细菌，完成这些 aims将提供重要的机制洞察到创新的对策，以打击寄生虫感染通过扰乱冬眠核糖体的生物发生和周转。