Characterization of the Cyclic Di-GMP Signaling System of Vibrio cholerae

描述(由申请人提供)：霍乱弧菌是世界卫生大流行霍乱的病原体，使用群体感应(QS)和环二GMP(c-diGMP)来调节生物膜的形成和毒力。QS是细菌分泌微小的化学信号来计算它们在种群中的数量并相应地控制群体行为的过程。C-diGMP是新近发现的第二信使细胞内信号分子，广泛存在于细菌中。我最近在霍乱弧菌中发现了qs和c-diGMP之间的一种新的联系。为了回答关于c-diGMP的一些基本问题，并进一步了解它与qs的联系，本研究提出：目的1.利用遗传筛选来鉴定c-diGMP与基因表达变化相关的转录成分；目的2.利用生物信息学和高通量化学筛选来鉴定霍乱弧菌中GGDEF/EAL蛋白的特定配体；以及目的3.利用X射线结晶学来确定GGDEF和EAL蛋白如何调节酶活性的信号识别。由于qs和c-diGMP都控制霍乱弧菌和许多其他细菌病原体的毒力，我希望这项工作可以导致治疗细菌性疾病的新的治疗方法。‘细菌感觉到多个微小的化学信号，以适应和响应他们的环境。这些化学信号通常控制细菌致病的能力。我对霍乱弧菌的研究试图揭开这些信号的本质，以及细菌如何对它们做出反应，以开发治疗细菌感染的新的治疗方法。相关性(参见说明)：K22研究学者发展奖将允许我在新的教师职位的头两年支持我的实验室成员(研究生、技术员和/或博士后)。这将允许我开发研究项目，这些项目将成为一项或多项R01拨款的基础。

DESCRIPTION (provided by applicant): Vibrio cholerae, the causative agent of the worldwide health pandemic cholera, uses both quorum sensing (QS) and cyclic di-GMP (c-diGMP) to regulate biofilm formation and virulence. QS is a process by which bacteria secrete small, chemical signals to count their numbers in a population and control group behavior accordingly. c-diGMP is a recently discovered second messenger intracellular signaling molecule ubiquitous in bacteria. I have recently discovered a novel connection between QS and c-diGMP in V. cholerae. To answer some of the fundamental question about c-diGMP, and further understand its connection to QS, this research proposes to: Aim 1. Identify the transcriptional components that couple c-diGMP to alterations in gene expression using genetic screens; Aim 2. Identify specific ligands of GGDEF/EAL proteins in V. cholerae using bioinformatics and a high-throughput chemical screen; and Aim 3. Utilize X-ray crystallography to determine how signal recognition by GGDEF and EAL proteins modulates enzymatic activity. As both QS and c-diGMP control virulence in V. cholerae and numerous other bacteria pathogens, it is my hope that this work could lead to novel therapeutic approaches for the treatment of bacterial disease. ' Bacteria sense multiple small chemical signals to adapt and respond to their environment. These chemical signals are often control the ability of bacteria to cause disease. My research in Vibrio cholerae seeks to unravel the nature of these signals, and how bacteria respond to them, to develop novel therapeutic approaches to treat bacterial infections. RELEVANCE (See instructions): The K22 Research Scholar Development Award will allow me to support members of my laboratory (graduate students, technician, and/or post-docs) during the first two years of my new faculty position. This will allow me to develop research projects that will form the basis of one or more R01 grants.