Regulation of Vibrio cholerae virulence by ToxR and TcpP

ToxR 和 TcpP 对霍乱弧菌毒力的调节

• 批准号: 7479790
• 负责人: ERIC S KRUKONIS
• 金额: $ 28.82万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-15 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7479790
• 关键词: Affect; Bile fluid; Binding; Binding Sites; Biochemical; Cells; Cessation of life; Cholera; Cholera Toxin; Condition; Cues; DNA Binding; DNA-Directed RNA Polymerase; Dehydration; Diarrhea; Disease; Elements; Ensure; Event; Fluorescence; Fluorescence Resonance Energy Transfer; Gene Expression; Genes; Genetic; Goals; Growth; In Vitro; Infection; Life; Localized; Measures; Mediating; Membrane; Numbers; Oral Rehydration Therapy; Pilum; Proteins; Regulation; Resistance; Role; System; Time; Toxin; Vibrio cholerae; Virulence; Virulence Factors; day; mutant; pandemic disease; porin; promoter; protein protein interaction; reconstitution; response; transcription factor

DESCRIPTION (provided by applicant): Vibrio cholerae is the causative agent of the diarrheal disease cholera, a debilitating disease affecting as many as ten million people annually. The disease is caused by cholera toxin, a protein whose expression is regulated by two membrane-localized transcription factors, ToxR and TcpP. This application investigates how ToxR and TcpP combine to activate the toxT promoter in V. cholerae, thus triggering virulence gene expression. In addition, we will investigate how ToxR activates a second promoter, ompLJ, in a TcpP- independent fashion. Expression of the porin, OmpU, leads to bile resistance in V. cholerae. We hypothesize that ToxR functions by different mechanisms at the ompU and toxT promoters. With ToxR directly activating ompU while serving to facilitate TcpP-mediated promoter activation at toxT. ' Finally, we will also develop a system for observing the the interaction of ToxR and TcpP in the membrane of living V. cholerae cells using FRET. This will allow us to observe a critic step in induction of virulence in \ real time as well as allow us to observe any changes in ToxR/TcpP interaction in response to a variety of environmental conditions. Specific Aims of this project are: I. Determine the mechanism of ompU and toxT activation by ToxR II. Determine the mechanisms of DNA-binding, ToxR interaction and RNA polymerase stimulation by TcpP and the role of each event in toxT activation III. Measure ToxR/TcpP protein-protein interactions in living V. cholerae cells after tcpP induction using FRET (fluorescence resonance energy transfer)

描述（由申请人提供）：Vibrio Cholerae是腹泻病霍乱的病因，这是一种使人衰弱的疾病，每年影响多达一千万人。该疾病是由霍乱毒素引起的，霍乱毒素是一种蛋白质，其表达受两个膜 - 定位转录因子ToxR和TCPP调节。该应用研究了如何组合TOXR和TCPP在V.霍乱中激活TOXT启动子，从而触发毒力基因表达。此外，我们将以TCPP独立的方式研究TOXR如何激活第二个启动子OMPLJ。孔蛋白的表达导致霍乱弧菌的胆汁耐药性。我们假设TOXR通过OMPU和TOXT启动子的不同机制发挥作用。 Toxr在用来促进TCPP介导的启动子激活的同时直接激活OPMU。 '最后，我们还将开发一个系统，用于观察使用FRET使用FRET的活体V.霍乱细胞膜中TOXR和TCPP的相互作用。这将使我们能够在实时诱导毒力方面观察批评者的一步，并允许我们观察到响应各种环境条件的TOXR/TCPP相互作用的任何变化。该项目的具体目的是：I。通过Toxr II确定OMPU和TOXT激活的机理。确定TCPP的DNA结合，TOXR相互作用和RNA聚合酶刺激的机制以及每个事件在TOXT激活III中的作用。使用FRET（荧光共振能传递）在TCPP诱导后，测量活体V.霍乱细胞中的TOXR/TCPP蛋白 - 蛋白质相互作用