Characterization of a novel ETEC virulence regulator

新型 ETEC 毒力调节剂的表征

• 批准号: 8091726
• 负责人: Joseph Thomas Wade
• 金额: $ 7万
• 依托单位: WADSWORTH CENTER
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2013-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8091726
• 关键词: Adhesions; Biochemical; Cessation of life; Child; DNA; Diarrhea; Epithelial Cells; Foundations; Future; Gene Expression Regulation; Gene Targeting; Genes; Genetic; Genomics; Goals; Human; Nucleotides; Plasmids; Play; Process; Proteins; RNA; Regulation; Resolution; Role; Site; Virulence; Work; enterotoxigenic Escherichia coli; genetic regulatory protein; insight; killings; novel; pathogenic bacteria; promoter; transcription factor

DESCRIPTION (provided by applicant): Gene regulation plays a critical role in the virulence of pathogenic bacteria. Our long-term goal is to understand how transcription factors (TFs) regulate virulence processes in the pathogenic species, Enterotoxigenic Escherichia coli (ETEC). ETEC is the leading bacterial cause of diarrhea in the developing world and causes >300,000 deaths annually, mainly in children. Many virulence genes have been identified in ETEC but relatively little is known about regulation of these genes. We have recently identified a TF, RoaA, encoded on one of the ETEC virulence plasmids, that is required for efficient adhesion to gut epithelial cells. We hypothesize that RoaA is a key regulator of ETEC virulence. The goal of this proposal is to comprehensively identify the regulatory targets of RoaA with nucleotide resolution. We will use complementary genomic approaches, ChIP-seq and RNA-seq, to identify direct and indirect regulatory targets. We will determine which of the target genes are required for ETEC virulence. We will then use genetic and biochemical analyses to identify the precise DNA sites for RoaA at the promoters of these genes. The proposed work will provide important insight into the regulation of ETEC virulence genes, and will form the foundation of future studies of ETEC virulence. PUBLIC HEALTH RELEVANCE: NARRATIVE Enterotoxigenic Escherichia coli (ETEC) kills more than 300,000 people annually. We have identified an ETEC regulatory protein that is required for efficient attachment to human gut epithelial cells. We will identify the regulatory targets of this protein.

描述（由申请人提供）： 基因调控在病原菌的毒力中起着至关重要的作用。我们的长期目标是了解转录因子 (TF) 如何调节致病物种产肠毒素大肠杆菌 (ETEC) 的毒力过程。 ETEC 是发展中国家腹泻的主要细菌原因，每年导致超过 300,000 人死亡，其中主要是儿童。已在 ETEC 中鉴定出许多毒力基因，但对这些基因的调控知之甚少。我们最近发现了一种 TF RoaA，它编码在一种 ETEC 毒力质粒上，它是有效粘附肠上皮细胞所必需的。我们假设 RoaA 是 ETEC 毒力的关键调节因子。该提案的目标是通过核苷酸分辨率全面识别 RoaA 的调控靶点。我们将使用互补的基因组方法、ChIP-seq 和 RNA-seq 来识别直接和间接的调控靶标。我们将确定 ETEC 毒力需要哪些靶基因。然后，我们将使用遗传和生化分析来确定这些基因启动子上 RoaA 的精确 DNA 位点。拟议的工作将为 ETEC 毒力基因的调控提供重要的见解，并将为未来 ETEC 毒力研究奠定基础。 公共卫生相关性： 叙述产肠毒素大肠杆菌 (ETEC) 每年导致超过 300,000 人死亡。我们已经鉴定出一种 ETEC 调节蛋白，它是有效附着于人肠道上皮细胞所需的。我们将确定该蛋白质的调控靶点。