Characterization of a novel family of Small Regulatory Proteins modulating virulence in Enterobacteriaceae

调节肠杆菌毒力的新型小调节蛋白家族的表征

• 批准号: 10560645
• 负责人: Araceli ELVIRA Santiago
• 金额: $ 41.12万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-02 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10560645
• 关键词: Acute; Affect; Animal Disease Models; Attenuated; Bacteria; Bacterial Genome; Binding; Biochemical; Biological Assay; Cells; Centers for Disease Control and Prevention (U.S.); ChIP-seq; Childhood; Citrobacter; Citrobacter rodentium; Clinical; Coupled; DNA Binding; DNA-Protein Interaction; Data; Developing Countries; Disease; Enterobacteriaceae; Epidemiology; Exhibits; Family; Fluorescence Microscopy; Gastroenteritis; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Transcription; Goals; Gram-Negative Bacteria; Histones; Homologous Gene; Hospitalization; Host Defense; Human; Impairment; In Vitro; Infection; Infection prevention; Intestines; Kidney; Knowledge; Lifting; Maps; Meningitis; Metabolism; Microscopy; Modeling; Molecular; Morbidity - disease rate; Pathogenesis; Phenotype; Play; Protein Family; Proteins; Public Health; Regulation; Regulator Genes; Regulon; Repression; Role; Salmonella; Shigella; Structure; Surface Plasmon Resonance; System; Therapeutic Intervention; Toxin; Type III Secretion System Pathway; Urinary tract infection; Vaccines; Vibrio; Virulence; Virulence Factors; Yersinia; biological adaptation to stress; clinically relevant; diarrheal disease; enteric infection; enteric pathogen; enteroaggregative Escherichia coli; enteropathogenic Escherichia coli; fitness; gene repression; genetic regulatory protein; genome-wide; gut colonization; gut inflammation; human stem cells; in vivo; insight; member; mortality; murine colitis; mutant; novel; novel therapeutics; novel vaccines; pathogen; pathogenic Escherichia coli; pathogenic bacteria; protein protein interaction; prototype; stem cell technology; stem cells; trait; transcriptome; vaccine strategy; virulence gene

PROJECT SUMMARY/ABSTRACT In the United States, the Center for Disease Control and Prevention (CDC) has estimated that more than 200 million episodes of acute gastroenteritis occur annually, resulting in nearly 1 million hospitalizations and over 5000 deceases. Vaccines against enteric pathogens are not available due, in part, to limited knowledge of the many virulence mechanisms that enteropathogens have evolved to colonize, multiply and escape host defenses. We have recently discovered the ANR (AraC Negative Regulators) family, comprising a large number of small regulatory proteins produced by diverse clinically important enteric pathogens including Vibrio spp., Salmonella spp., Shigella spp., Yersinia spp., Citrobacter spp., enterotoxigenic (ETEC), enteropathogenic EPEC, enterohaemorragic (EHEC) and enteroaggregative E. coli (EAEC). Members of the ANR family modulate the expression of virulence factors such as fimbriae, toxins, type-3 and type-6 secretion systems, and genes associated with metabolism, stress-response and fitness, by protein-protein interactions with positive transcriptional regulators of the AraC/XylS family and negative regulators of the HNS family. Furthermore, the absence of this regulatory system affects bacterial pathogenesis in a natural model of infection. Our overall hypothesis is that the ANR family plays a critical role in the concerted regulation of fitness and virulence in pathogens of the Enterobacteriaceae family by taking concomitant control of AraC and HNS global regulators. Here, we propose to obtain mechanistic insights into ANR molecular interactions (AIM-1), regulation (AIM 2) and pathogenesis (AIM-3) using EAEC, EPEC and C. rodentium as pathogen models, human stem cell technology and a natural animal model of disease, coupled with genetic, biochemical and molecular approaches such as Surface Plasmon Resonance (SPR), Time-lapse fluorescence microscopy (TLFM) and ChIP-seq. Our proposed studies will advance our knowledge of global gene regulatory mechanisms that govern fitness and virulence in pathogenic bacteria, and will likely open new avenues for therapeutic intervention.

项目概要/摘要 在美国，疾病控制与预防中心 (CDC) 估计有超过 200 每年发生 100 万例急性胃肠炎，导致近 100 万人住院治疗 5000人死亡。无法获得针对肠道病原体的疫苗，部分原因是对肠道病原体的了解有限。 肠道病原体已经进化出许多毒力机制来定植、繁殖和逃离宿主 防御。我们最近发现了 ANR（AraC 负调节因子）家族，包括一个大的 由多种临床重要的肠道病原体（包括弧菌）产生的小调节蛋白的数量 spp.、沙门氏菌属、志贺氏菌属、耶尔森氏菌属、柠檬酸杆菌属、产肠毒素 (ETEC)、肠致病性 EPEC、肠出血性大肠杆菌 (EHEC) 和肠聚集性大肠杆菌 (EAEC)。 ANR家族成员 调节毒力因子的表达，例如菌毛、毒素、3 型和 6 型分泌系统， 以及与新陈代谢、压力反应和健康相关的基因，通过蛋白质-蛋白质相互作用 AraC/XylS 家族的正转录调节因子和 HNS 家族的负转录调节因子。 此外，这种调节系统的缺失会影响自然模型中的细菌发病机制。 感染。我们的总体假设是 ANR 家族在健康的协调调节中发挥着关键作用 通过同时控制 AraC 和 HNS 来控制肠杆菌科病原体的毒力 全球监管机构。在这里，我们建议获得 ANR 分子相互作用（AIM-1）的机制见解， 使用 EAEC、EPEC 和 C. rodentium 作为病原体模型进行调节（AIM 2）和发病机制（AIM-3）， 人类干细胞技术和自然疾病动物模型，加上遗传、生化和 分子方法，例如表面等离子共振 (SPR)、延时荧光显微镜 (TLFM) 和 ChIP-seq。我们提出的研究将增进我们对全球基因调控的了解 控制病原菌适应性和毒力的机制，可能会开辟新的途径 治疗干预。