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.

抽象的 致病性微生物的代谢必须是柔性的，因为病原细菌的有效复制 主机取决于主动适应过程。细菌病原体弧菌霍乱的人类感染需要 多种适应过程最终导致小肠中强大的毒力因子产生。虽然 在几种病原体中已经描述了代谢状态和毒力因子表达之间的联系，尚未 在V.霍乱中进行了详细研究。同样，膜生物能力在细菌毒力中的作用不好 理解，尽管大多数病原生物都广泛利用膜呼吸作为其一部分生理学。我们的 先前的结果表明，膜生物能学的变化会影响霍乱弧菌中的毒力基因表达。更多的 具体而言，主要V. Cholerae呼吸酶（NQR）的丧失导致毒力基因转录的变化。这里， 我们建议对V中的膜呼吸与毒力基因调节之间的联系进行研究。 霍乱，重点是活性氧（ROS）和喹酮池。已经有很好的记录 多种毒力相关的转录因子通过翻译后修饰在霍乱弧菌中感知氧化应激 例如硫醇开关。我们以前已经建立了NQR作为周质和细胞质ROS的来源。 此外，我们最近开发了一种通过操纵表达来产生V.霍乱弧菌的ROS的方法 NQR和并行的水平，可以使用新型的基于荧光团的活细胞中的细胞质ROS水平来监测 方法。该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相关联 V.霍乱的因素。总体而言，我们希望能够获得对这种生物本身的代谢能力的重要见解和 它与毒力基因表达的关系是发病机理的重要方面。我们对该项目的理由是 生成有关V.霍乱代谢及其与毒力因子产生的联系的基本信息将 为感染的新型治疗策略提供了潜在的机会。