FimX in Regulation of Type IV Pili Expression

FimX 对 IV 型菌毛表达的调节

• 批准号: 6886645
• 负责人: THOMAS SCOT MURRAY
• 金额: $ 5.35万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6886645
• 关键词: Pseudomonas aeruginosa; adenosinetriphosphatase; binding sites; cell motility; chemical kinetics; enzyme activity; guanosine triphosphate; guanylate cyclase; immunoprecipitation; membrane transport proteins; molecular assembly /self assembly; pilin; postdoctoral investigator; protein binding; protein protein interaction; protein quantitation /detection; second messengers; transcription factor; virulence; yeast two hybrid system

DESCRIPTION (provided by applicant): FimX is a recently described protein in Pseudomonas aeruginosa (PA) shown to regulate type IV pill (TFP), an important virulence factor, in response to environmental signals. We propose to characterize the role of FimX, a GGDEF/EAL containing protein, in TFP assembly. We will confirm that FimX synthesizes cyclic di-GMP (c-diGMP), a regulatory molecule hypothesized to be important for a wide variety of cellular functions. The contributions of each FimX domain to regulation of c-diGMP will be elucidated. We have identified two components of ABC transporters that partner with FimX in a yeast two-hybrid screen. We will confirm these interactions with co-immunoprecipitation studies and identify the FimX domains required to bind these proteins. Furthermore, we will generate PA strains lacking these ABC transporters and confirm their role in the regulation of TFP. Additional studies will determine if these transporters are exporters or importers. These studies will determine whether c-diGMP is a second messenger regulating twitching motility in PA. We also will characterize the first ABC transporters identified in PA involved in controlling TFP expression.

描述（由申请人提供）：FIMX是铜绿假单胞菌（PA）中最近描述的蛋白质，用于调节对环境信号的响应，以调节IV型药丸（TFP），这是一个重要的毒力因子。我们建议表征TFP组装中含有GGDEF/EAL的FIMX的作用。我们将确认FIMX合成环状DI-GMP（C-DIGMP），这是一种对多种细胞功能很重要的调节分子。每个FIMX结构域对C-二烯的调节的贡献将得到阐明。我们已经确定了ABC转运蛋白的两个组件，它们与酵母双杂交屏幕中的FIMX合作。我们将确认与共免疫沉淀研究的这些相互作用，并确定结合这些蛋白质所需的FIMX结构域。此外，我们将产生缺乏这些ABC转运蛋白的PA菌株，并确认其在TFP调节中的作用。其他研究将确定这些转运蛋白是出口商还是进口商。这些研究将确定C-DIGMP是否是PA中的第二个Messenger调节抽动运动。我们还将表征参与控制TFP表达的PA中鉴定出的第一个ABC转运蛋白。