Quantitative analysis of growth in a streamlined obligate intracellular pathogen

流线型专性细胞内病原体生长的定量分析

• 批准号: 10361525
• 负责人: Erin D Goley
• 金额: $ 24.56万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-01 至 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10361525
• 关键词: Actins; Address; Animals; Anti-Bacterial Agents; Antibiotics; Bacteria; Bioinformatics; Biology; Cell Cycle; Cell Cycle Progression; Cell Shape; Cell Wall; Cells; Chemicals; Complex; Cytolysis; Cytoplasm; Defect; Development; Disease; Environment; Enzymes; Escherichia coli; Evolution; Fluorescent Probes; Foundations; Genome; Growth; Homologous Gene; Human; Image Analysis; Individual; Kinetics; Knowledge; Life; Life Style; Link; Measures; Metabolic; Metabolic Pathway; Metabolism; Methods; Microscopy; Modeling; Modification; Monitor; Morphogenesis; Morphology; Nature; Organism; Pathogenesis; Pathway interactions; Pattern; Peptides; Peptidoglycan; Pharmacology; Polymers; Preventive; Process; Proteins; Regulation; Reporting; Resolution; Rickettsia; Rickettsia Infections; Rickettsia parkeri; Rod; Role; Severities; Shapes; Spatial Distribution; Therapeutic; Tick-Borne Diseases; Work; crosslink; human pathogen; imaging modality; member; pathogen; pathogenic bacteria; protein distribution; small molecule inhibitor; spotted fever; sugar; synthetic enzyme; tick-borne; time use; tool

Members of the Spotted Fever Group (SFG) of the bacterial genus Rickettsia are obligate intracellular pathogens that cause spotted fever disease in humans ranging from mild to life-threatening. Like all species in the order Rickettsiales, SFG bacteria have undergone genome streamlining and are dependent on a eukaryotic host for dozens of essential metabolites. Notably, reductive evolution has led to loss or modification of factors in metabolic and morphogenetic pathways predicted to impact peptidoglycan cell wall metabolism. Peptidoglycan metabolism is essential for growth and viability and is therefore an effective antibiotic target. The impact of genome streamlining on the mechanisms of growth and division of the Rickettsiales, as compared to their free- living relatives, is not clear. This gap in knowledge persists, in part, because we lack tools to investigate growth of intracellular bacteria with sufficient resolution to derive a mechanistic understanding. We hypothesize that reductive evolution of metabolic and morphogenetic pathways necessitates diversification of known mechanisms to support intracellular growth. In this proposal, we aim to develop and apply methods to quantitatively analyze growth and cell wall metabolism of Rickettsia parkeri in eukaryotic host cells. R. parkeri is a SFG organism that causes relatively mild disease. As such, it is a BSL2 pathogen that has been studied as a representative to understand SFG pathogenesis. Through the proposed work, we will expand the study of R. parkeri to investigate its intracellular growth mechanisms. In aim 1, we will develop and apply imaging methods to quantitatively analyze the cell shape of, subcellular protein distribution in, cell cycle status of, and kinetics of growth of individual R. parkeri in the cytoplasm of eukaryotic host cells. In aim 2, we will determine the chemical composition of and spatial patterning of peptidoglycan cell wall metabolism of R. parkeri growing in the host cytoplasm. Completion of the proposed aims will provide foundational knowledge on the growth kinetics and mechanisms of PG metabolism of a tick-borne, obligate intracellular human pathogen and establish broadly applicable quantitative approaches for studying growth of bacteria within a eukaryotic host cell. Ultimately, the information and methods derived from this project will inform development of preventive and therapeutic strategies for limiting Rickettsial disease.

细菌属的斑点发烧组（SFG）的成员是强性细胞内病原体 这会导致人类发现的发烧疾病，从轻度到威胁生命。像所有物种一样 立克菌，SFG细菌经历了基因组的精简，并取决于真核宿主 数十个基本代谢产物。值得注意的是，还原性演变导致因素的损失或修改 预测将影响肽聚糖细胞壁代谢的代谢和形态发生途径。肽聚糖 代谢对于生长和生存能力至关重要，因此是一个有效的抗生素靶标。的影响 基因组简化了立克人士的生长和分裂的机制，与他们的自由相比 亲戚不清楚。知识差距仍然存在，部分原因是我们缺乏研究增长的工具 具有足够分辨率的细胞内细菌，可以得出机理的理解。我们假设这一点 代谢和形态发生途径的还原性演变需要多样化已知机制 支持细胞内生长。在此提案中，我们旨在开发和应用方法来定量分析 真核宿主细胞中立克帕克里的生长和细胞壁代谢。 R. Parkeri是一种SFG生物体 引起相对温和的疾病。因此，它是一种已被研究为代表的BSL2病原体 了解SFG发病机理。通过拟议的工作，我们将扩大对R. Parkeri的研究以调查 其细胞内生长机制。在AIM 1中，我们将开发并应用成像方法来定量 分析个体生长的细胞形状，亚细胞蛋白分布，细胞周期状态和动力学 真核宿主细胞的细胞质中的R. parkeri。在AIM 2中，我们将确定和 在宿主细胞质中生长的R. parkeri的肽聚糖细胞壁代谢的空间模式。完成 拟议的目标将提供有关PG生长动力学和机制的基本知识 tick传播的代谢，义务细胞内人体病原体并建立广泛适用的定量 研究真核宿主细胞中细菌生长的方法。最终，信息和方法 从该项目中得出的将为您的预防和治疗策略的发展提供限制的限制立体的策略 疾病。

项目成果

Quantitative analysis of morphogenesis and growth dynamics in an obligate intracellular bacterium.

• DOI: 10.1091/mbc.e23-01-0023
• 发表时间: 2023-06-01
• 期刊: MOLECULAR BIOLOGY OF THE CELL
• 影响因子: 3.3
• 作者: Figueroa-Cuilan, Wanda M.;Irazoki, Oihane;Feeley, Marissa;Smith, Erika;Nguyen, Trung;Cava, Felipe;Goley, Erin D.
• 通讯作者: Goley, Erin D.