The role of NQR in ROS-dependent virulence regulation in Vibrio cholerae

NQR 在霍乱弧菌 ROS 依赖性毒力调节中的作用

• 批准号: 10721326
• 负责人: Claudia C Hase
• 金额: $ 7.43万
• 依托单位: OREGON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-08 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10721326
• 关键词: 3-Dimensional; Address; Affect; Antioxidants; Bacteria; Bacterial Infections; Bacterial Physiology; Biochemical; Bioenergetics; Biological Assay; Cells; Communicable Diseases; Complex; Cytoplasm; Development; Devices; Electron Transport; Electron Transport Complex III; Ensure; Environment; Enzymes; Exposure to; Future; Gastrointestinal tract structure; Gene Expression; Gene Expression Regulation; Generations; Genetic Transcription; Goals; Growth; Human; Hydroquinones; Infection; Investigation; Link; Membrane; Metabolic; Metabolism; Methods; Modeling; Molecular; Monitor; NADH; Na(+)-K(+)-Exchanging ATPase; Organism; Oxidases; Oxidation-Reduction; Oxidative Stress; Oxidoreductase; Oxygen; Pathogenesis; Pathogenicity; Pathway interactions; Physiology; Play; Post-Translational Protein Processing; Process; Production; Quinones; Reactive Oxygen Species; Regulation; Reporting; Research; Respiration; Role; Signal Induction; Signal Transduction; Site; Site-Directed Mutagenesis; Small Intestines; Solid; Source; Stress; Sulfhydryl Compounds; Superoxides; System; Testing; Therapeutic Intervention; Vibrio cholerae; Virulence; Virulence Factors; Work; bacterial metabolism; complex IV; experimental study; extracellular; flexibility; fluorophore; in vivo; insight; microbial; microorganism; mutant; novel; pathogen; pathogenic bacteria; pathogenic microbe; periplasm; respiratory; response; three dimensional structure; tool; transcription factor; treatment strategy; ubiquinol; ubisemiquinone; virulence gene

ABSTRACT The metabolism of pathogenic microorganisms has to be flexible as the efficient replication of pathogenic bacteria in the host depends on an active adaptation process. Human infection by the bacterial pathogen Vibrio cholerae, requires multiple adaptation processes that ultimately lead to robust virulence factor production in the small intestine. Although a link between the metabolic status and virulence factor expression has been described in several pathogens, it has not been investigated in detail in V. cholerae. Similarly, the role of membrane bioenergetics in bacterial virulence is not well understood, although most pathogenic organisms extensively utilize membrane respiration for part of their physiology. Our previous results suggest that changes in membrane bioenergetics affect virulence gene expression in V. cholerae. More specifically, loss of the main V. cholerae respiration enzyme (NQR) leads to changes in virulence gene transcription. Here, we propose to perform an investigation into the link between membrane respiration and virulence gene regulation in V. cholerae, with an emphasis on reactive oxygen species (ROS) and the quinone pool. It has been well documented that multiple virulence-associated transcription factors sense oxidative stress in V. cholerae via post-translational modifications such as thiol-switches. We have previously established NQR as a source of both periplasmic as well as cytoplasmic ROS. Moreover, we recently developed a method to produce varying levels of ROS in V. cholerae by manipulating the expression level of NQR and in parallel, the level of cytoplasmic ROS in live cells can be monitored using a novel, fluorophore-based approach. This system enables us to quantitatively correlate ROS levels with the expression of genes encoding for virulence factors in V. cholerae. Overall, we expect to gain important insights into the metabolic capacity of this organism per se and its relation to virulence gene expression as an important aspect of pathogenesis. Our rationale for this project is that the generation of fundamental information concerning V. cholerae metabolism and its link to virulence factor production will provide potential opportunities for novel treatment strategies for infections.

摘要 病原微生物的新陈代谢必须是灵活的，因为病原细菌在 东道主依赖于一个积极的适应过程。人类感染霍乱弧菌，需要 多个适应过程，最终导致在小肠产生强大的毒力因子。尽管一个 新陈代谢状态和毒力因子表达之间的联系已经在几种病原体中被描述，但还没有 对霍乱弧菌进行了详细的调查。同样，膜生物能量学在细菌毒力中的作用也不是很好。 理解，尽管大多数病原体广泛地利用膜呼吸作为其生理的一部分。我们的 以往的研究结果表明，膜生物能量学的改变影响霍乱弧菌毒力基因的表达。更多 具体地说，霍乱弧菌主要呼吸酶(NQR)的丧失会导致毒力基因转录的变化。这里, 我们建议对膜呼吸和毒力基因调控之间的联系进行研究。 霍乱弧菌，重点是活性氧物种(ROS)和苯二酚池。已经有很好的证据表明 多个毒力相关转录因子通过翻译后修饰感知霍乱弧菌的氧化应激 例如硫醇开关。我们以前已经确定NQR既是周质ROS的来源，也是细胞质ROS的来源。 此外，我们最近开发了一种方法，通过操纵表达式在霍乱弧菌中产生不同水平的ROS NQR水平，同时，活细胞中细胞质ROS水平可以使用一种新型的基于荧光团的监测 接近。这个系统使我们能够定量地将ROS水平与编码毒力的基因的表达联系起来 霍乱弧菌的致病因素。总体而言，我们希望对这种有机体本身的新陈代谢能力有重要的了解 其与毒力基因表达的关系作为致病机理的一个重要方面。我们这个项目的理由是 关于霍乱弧菌代谢及其与毒力因子产生的联系的基本信息的产生将 为新的感染治疗策略提供潜在的机会。