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

基本信息

批准号：
8632606
负责人：
STUART A THOMPSON
金额：
$ 33.7万
依托单位：
AUGUSTA UNIVERSITY
依托单位国家：
美国
项目类别：
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/8632606
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 Food Processing Freezing Gastroenteritis Gene Expression Gene Expression Regulation Genetic Goals Gram-Negative Bacteria Growth 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 Reactive Arthritis Refrigeration Regulation Regulon Relative (related person)Resistance Role Route Salmonella Shigella Sigma Factor Signaling Molecule Small RNA Structure Syndrome System Time Translations United States Virulence Virulence Factors Water Work cell motility extracellular foodborne member mutant novel protein expression protein protein interaction public health relevance quorum sensing response wild bird

项目摘要

ABSTRACT Campylobacter jejuni is a primary bacterial cause of gastroenteritis in the United States, with at least 2 million cases of C. jejuni gastroenteritis each 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-Barr¿ 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.

抽象的弯曲杆菌空肠是美国胃炎的主要原因，至少有2个美国每年在美国每年有百万杆菌肠胃炎病例（这一事件等于或更大沙门氏菌和志贺氏菌组合）。 C. jejuni负责零星疾病以及食物传播 /水 - 天生的爆发是由污染的食物和娱乐水域的暴露造成的。一些C. jejuni 感染导致Guillain-Barr¿综合征的发展，这是急性瘫痪的主要原因世界。尽管弯曲杆菌疾病的患病率很高，并且研究了20多年，但旋转梭菌引起疾病的机制仍然不完全理解并被严重理解。在许多细菌中，CSRA调节许多重要的表型，包括病毒，碳代谢，运动性，法定感测，生物膜产生和动物定植。而在所有革兰氏阴性细菌中研究到迄今为止，CSRA活动由诱导的小调节RNA（SRNA）控制，新数据建议首次与FLIW的新型蛋白质 - 蛋白质相互作用来调节J. jejuni CSRA，这是一个成员鞭毛生物合成途径，因此将关键病毒因子鞭毛的合成与调节其他固定相细胞过程，例如生物膜形成。我们构建了一个C. jejuni 缺乏CSRA的突变体，并表明CSRA突变体表现出与多效性病毒相关的表型包括运动性降低，上皮细胞依从性，生物膜形成，对氧化应激的耐药性和小鼠的定殖。相反，CSRA突变体显示出人类上皮细胞的侵袭增加。这强调了CSRA调节型在空肠梭菌发病机理中的重要性。我们已经确定了假定的CSRA调节，它包含许多蛋白质以及固定相过程，例如生物膜形成和乙酸盐代谢。最后，我们有证明了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：空肠C. c. jejuni csra在固定的固定杆菌的发病机理中起重要作用阶段，病毒 /生存特性的转录后调节，包括运动性和生物膜形成。我们提出了一项关于CSRA介导的转录后固定相基因调控的详细研究。空肠，重点是CSRA控制运动性和生物膜形成的机制。我们将使用遗传，蛋白质组学和生化方法来实现这两个特定的目标目的：目标1）定义CSRA，FLIW和FLAA在协调固定相调节运动和趋化性和目标2）定义CSRA在控制生物膜生产和乙酸盐代谢中的作用。