Coordinated stationary phase control of Campylobacter motility and biofilm

弯曲杆菌运动和生物膜的协调固定相控制

• 批准号: 8967554
• 负责人: STUART A THOMPSON
• 金额: $ 30万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-12-01 至 2018-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8967554
• 关键词: Acetates; Acute; Address; Adherence; Anabolism; Animals; Bacillus subtilis; Bacteria; Bacteria sigma factor KatF protein; Binding Proteins; Biochemical; Campylobacter; Campylobacter infection; Campylobacter jejuni; Carbon; Cattle; Cell physiology; Cells; Cellular Structures; Characteristics; Chemotaxis; Competitive Binding; Consumption; Data; Development; Disease; Disease Outbreaks; Domestic Fowls; Enteral; Epithelial Cells; Escherichia coli; Exhibits; Exposure to; Flagella; Food Contamination; Food Processing; Freezing; Gastroenteritis; Gene Expression; Gene Expression Regulation; Genetic; Goals; Gram-Negative Bacteria; Growth; Guillain-Barré Syndrome; Health; High Prevalence; Human; Incidence; Infection; Ingestion; Lead; Link; Meat; Mediating; Messenger RNA; Metabolism; Microbial Biofilms; Mus; Nature; Oxidative Stress; Paralysed; Pathogenesis; Pathway interactions; Persons; Phase; Phenotype; Physiological; Play; Polysaccharides; Post-Transcriptional Regulation; Process; Production; Property; Proteins; Proteomics; Refrigeration; Regulation; Regulon; Reiter Disease; Resistance; Role; Route; Salmonella; Shigella; Sigma Factor; Signaling Molecule; Small RNA; Structure; System; Time; Translations; United States; Virulence; Virulence Factors; Water; Work; cell motility; extracellular; foodborne; logarithm; member; mutant; novel; protein expression; protein protein interaction; quorum sensing; response; wild bird

DESCRIPTION (provided by applicant): Campylobacter jejuni is a primary bacterial cause of gastroenteritis in the United States, with at least 2 million cases of C. jejuni gastroenteritis eah year in the U.S. (an incidence equal or greater to that of Salmonella and Shigella combined). C. jejuni is responsible for sporadic disease as well as food-borne / water- borne outbreaks, which result from contaminated food and exposure to recreational waters. Some C. jejuni infections lead to the development of Guillain-Barré Syndrome, the leading cause of acute paralysis in the world. Despite the high prevalence of Campylobacter disease and more than 20 years of study, the mechanisms by which C. jejuni causes disease remain incompletely understood and severely understudied. In many bacteria, CsrA regulates numerous important phenotypes, including virulence, carbon metabolism, motility, quorum sensing, biofilm production, and animal colonization. While in all Gram-negative bacteria studied to date CsrA activity is controlled by inducible small regulatory RNAs (sRNAs), new data suggests for the first time that C. jejuni CsrA is regulated by novel protein-protein interactions with FliW, a member of the flagellar biosynthesis pathway, thus linking the synthesis of the critical virulence factors flagella to the regulation of other stationary phase cell processes such as biofilm formation. We constructed a C. jejuni mutant lacking csrA, and have shown that the csrA mutant exhibits pleiotropic virulence-related phenotypes including decreased motility, epithelial cell adherence, biofilm formation, resistance to oxidative stress, and colonization of mice. Conversely, the csrA mutant shows increased invasion of human epithelial cells. This underscores the importance of the CsrA regulon in C. jejuni pathogenesis. We have determined the presumptive CsrA regulon, and it contains a number of proteins with clear links to flagellar motility / chemotaxis and to stationary phase processes such as biofilm formation and acetate metabolism. Finally, we have demonstrated direct interaction of FliW with CsrA, supporting the link between flagella and CsrA regulation. Overall hypothesis: C. jejuni CsrA plays an important role in the pathogenesis of C. jejuni via stationary phase, post-transcriptional regulation of virulence / survival properties including motility and biofilm formation. We propose a detailed study of CsrA-mediated post-transcriptional stationary phase gene regulation in C. jejuni, focusing on the mechanism by which CsrA controls the expression of motility and biofilm formation. We will use genetic, proteomic and biochemical approaches to achieve the goals outlined in these two specific aims: Aim 1) Define the roles of CsrA, FliW, and FlaA in coordinating stationary phase regulation of motility and chemotaxis, and Aim 2) Define the role of CsrA in regulating biofilm production and acetate metabolism.

描述（由申请方提供）：空肠弯曲杆菌是美国胃肠炎的主要细菌病因，至少有200万例空肠弯曲杆菌病例。在美国，空肠胃肠炎每年发生一次（发病率等于或大于沙门氏菌和志贺氏菌的总和）。C.空肠是散发性疾病以及食源性/水源性疾病爆发的原因，这些疾病爆发是由受污染的食物和接触娱乐沃茨引起的。一些C。空肠感染导致格林-巴利综合征（Guillain-Barré Syndrome）的发展，格林-巴利综合征是世界上急性瘫痪的主要原因。尽管弯曲杆菌病的高患病率和20多年的研究，C。空肠引起的疾病仍然不完全了解和严重不足的研究。 在许多细菌中，CsrA调节许多重要的表型，包括毒力、碳代谢、运动性、群体感应、生物膜产生和动物定殖。虽然在迄今为止研究的所有革兰氏阴性细菌中，CsrA活性都是由可诱导的小调控RNA（sRNA）控制的，但新的数据首次表明，C。空肠CsrA通过与鞭毛生物合成途径的成员FliW的新型蛋白质-蛋白质相互作用来调节，从而将关键毒力因子鞭毛的合成与其他稳定期细胞过程（例如生物膜形成）的调节相联系。我们构建了一个C。空肠突变体缺乏csrA，并已显示csrA突变体表现出多效性毒性相关表型，包括运动性降低、上皮细胞粘附、生物膜形成、对氧化应激的抗性和小鼠定殖。相反，csrA突变体显示出增加的人上皮细胞侵袭。这强调了CsrA调节子在C.空肠发病机制我们已经确定了假定的CsrA调节子，它包含了一些蛋白质与鞭毛运动/趋化性和固定相过程，如生物膜形成和乙酸盐代谢有明确的联系。最后，我们已经证明了FliW与CsrA的直接相互作用，支持鞭毛和CsrA调节之间的联系。 总体假设：C.空肠CsrA在C.空肠通过稳定期，转录后调节毒力/存活特性，包括运动性和生物膜形成。 我们提出了一个详细的研究环孢素A介导的转录后稳定期基因调控在C。空肠，侧重于CsrA控制运动和生物膜形成的表达的机制。我们将使用遗传学，蛋白质组学和生物化学方法来实现这两个特定目标中概述的目标：目的1）定义CsrA，FliW和FlaA在协调运动性和趋化性的稳定期调节中的作用，目的2）定义CsrA在调节生物膜产生和乙酸盐代谢中的作用。