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.

立克次体属斑点热病群 (SFG) 的成员是专性细胞内病原体 引起人类斑点热病的疾病从轻微到危及生命。与该目中的所有物种一样 立克次体、SFG 细菌已经经历了基因组精简，并且依赖于真核宿主 数十种必需代谢物。值得注意的是，还原进化导致了一些因素的丢失或修改。 代谢和形态发生途径预计会影响肽聚糖细胞壁代谢。肽聚糖 新陈代谢对于生长和生存至关重要，因此是有效的抗生素靶点。的影响 与其自由生长和分裂机制相比，基因组精简对立克次氏体的生长和分裂机制的影响 活着的亲戚，不清楚。这种知识差距持续存在，部分原因是我们缺乏调查增长的工具 以足够的分辨率对细胞内细菌进行观察，以获得机制上的理解。我们假设 代谢和形态发生途径的还原进化需要已知机制的多样化 以支持细胞内生长。在本提案中，我们的目标是开发和应用定量分析的方法 帕克立克次体在真核宿主细胞中的生长和细胞壁代谢。 R. Parkeri 是一种 SFG 生物体， 引起相对轻微的疾病。因此，它是一种 BSL2 病原体，已作为代表进行了研究 了解 SFG 发病机制。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。在目标 1 中，我们将开发并应用成像方法来定量 分析个体的细胞形状、亚细胞蛋白质分布、细胞周期状态和生长动力学 真核宿主细胞细胞质中的帕克里氏菌。在目标 2 中，我们将确定 和 的化学成分 在宿主细胞质中生长的 R. parkeri 肽聚糖细胞壁代谢的空间模式。完成 所提出的目标将提供有关 PG 生长动力学和机制的基础知识 蜱传的、专性细胞内人类病原体的代谢，并建立广泛适用的定量 研究真核宿主细胞内细菌生长的方法。最后，信息和方法 该项目的成果将为限制立克次体的预防和治疗策略的制定提供信息 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.