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

基本信息

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
项目类别：
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

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

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项目摘要

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.

项目摘要/摘要在美国，疾病控制与预防中心（CDC）估计有200多个每年发生数百万次急性胃肠炎，导致近100万个住院 5000次。针对肠道病原体的疫苗不可用，部分原因是对肠病原体的许多毒力机制已进化为殖民，繁殖和逃生宿主防御。我们最近发现了ANR（ARAC负调节器）家族，包括一个大型由临床上重要的肠道病原体产生的小调节蛋白的数量，包括颤音 Spp。，沙门氏菌属，志贺氏菌，耶尔森氏菌，柠檬酸菌，肠毒素（ETEC），肠病毒 EPEC，EnteroHaeMorragic（EHEC）和肠肠肠球菌（EAEC）。 ANR家族的成员调节毒力因子的表达，例如fimbriae，毒素，3型和6型分泌系统，以及与蛋白质 - 蛋白质相互作用与代谢，应激反应和适应性相关的基因 HNS家族的ARAC/Xyls家族和负调节剂的阳性转录调节剂。此外，这种调节系统的不存在会影响细菌发病机理感染。我们的总体假设是，ANR家族在适应性的一致调节中起着至关重要的作用通过同时控制ARAC和HNS，在肠杆菌科家族的病原体中毒力全球监管机构。在这里，我们建议获得对ANR分子相互作用（AIM-1）的机械见解，使用EAEC，EPEC和啮齿动物C.作为病原体模型的调节（AIM 2）和发病机理（AIM-3），人类干细胞技术和自然动物模型，再加上遗传，生化和分子方法，例如表面等离子体共振（SPR），延时荧光显微镜（TLFM）和Chip-Seq。我们提出的研究将提高我们对全球基因调节的了解控制致病细菌的适应性和毒力的机制，可能会为治疗干预。