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：
10522119
负责人：
Mee-Ngan F Yap
金额：
$ 33.69万
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/10522119
关键词：
Address Area Bacteria Binding Biochemistry Biogenesis Biological Catalysis Cell Survival Cells Code Comparative Study Complex Conflict (Psychology)Crystallization Cytoplasmic Protein Data Development Dimerization Endoribonucleases Escherichia coli Eukaryota Exonuclease Exoribonucleases Firmicutes Genetic Screening Genetic Translation Goals Gram-Negative Bacteria Gram-Positive Bacteria Growth Guanosine Triphosphate Heterogeneity Hibernation Hydrolysis Infection Intervention Knock-out Knowledge Life Link Location Longevity Mass Spectrum Analysis 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 Process Translational Derepression Translations Virulence Factors Work antimicrobial bacterial genetics cell growth chronic infection combat dimer 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rapid growth ribonuclease R segregation small molecule inhibitor spatiotemporal structural biology translation factor unpublished works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 In our unpublished work, we isolated an additional ribonuclease mutant that rescues the loss of ribosomes in S. aureus.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都使用的保守机制来防止翻译并延长有机寿命。来自各种细菌种类的最新研究，包括致病性金黄色葡萄球菌为冬眠100s核糖体的关键作用提供了令人信服的证据除了阻止翻译计划外，还要保护核糖体池免受损伤。我们发现金黄色葡萄球菌缺乏休眠促进系数（HPF）的核糖体迅速被3'-5'Exoniclease rnase降解 R和其他迄今未知的核糖珠。在未发表的工作中，我们隔离了额外的核糖核酸酶挽救金黄色葡萄球菌核糖体损失的突变体。令人惊讶的是，我们发现核糖体不是唯一的金黄色葡萄球菌HPF的靶标；相反，HPF可以与未知生物活性的细胞质蛋白相互作用，从而减少了冬眠100s核糖体的抽象。我们进一步证明了HPF受到限制在快速增长过程中，特定的亚细胞定位，可罕见地瞥见可能的HPF隔离通过积极翻译核糖体。在此提案中，我们将采用高度多学科的方法由结构生物学，法学，细菌遗传学，生物化学和高分辨率显微镜组成实现以下目标：（1）确定HPF保护核糖体免受的分子机制核糖核解裂解。（2）确定HPF先前未被发现的外聚体作用。（3）确定 HPF的时空定位如何避免翻译冲突。 HPF和RNase R是进化医院革兰氏阳性和革兰氏阴性细菌中保守的病毒因子，这些因素的完成目的将提供对创新反策略的重要机械洞察力，以打击顽固感染通过扰动冬眠核糖体的生物发生和周转。