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）： 基因调节在致病细菌的毒力中起关键作用。我们的长期目标是了解转录因子（TFS）如何调节致病性物种，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原因，每年造成300,000人死亡，主要是儿童。在ETEC中已经鉴定出许多毒力基因，但对这些基因的调节知之甚少。我们最近已经确定了在一个ETEC毒力质粒之一上编码的TF ROAA，这是对肠道上皮细胞有效粘附所必需的。我们假设ROAA是ETEC毒力的关键调节剂。该提案的目的是通过核苷酸分辨率全面确定ROAA的调节靶标。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节目标。我们将确定靶基因的ETEC毒力需要哪些。然后，我们将使用遗传和生化分析来识别这些基因启动子的ROAA的精确DNA位点。拟议的工作将为ETEC毒力基因的调节提供重要的见解，并将构成对ETEC毒力的未来研究的基础。 公共卫生相关性： 叙事肠毒素大肠杆菌（ETEC）每年杀死30万以上的人。我们已经确定了有效附着在人肠上皮细胞上所需的ETEC调节蛋白。我们将确定该蛋白质的调节靶标。