Chemical genetic approach to Vibrio cholerae pathogensis

霍乱弧菌致病的化学遗传学方法

• 批准号: 6931448
• 负责人: DEBORAH T HUNG
• 金额: $ 12.53万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2008-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6931448
• 关键词: Vibrio cholerae; bacteria infection mechanism; bacterial genetics; bacterial proteins; bioterrorism /chemical warfare; chemical genetics; cholera; cholera toxin; expression cloning; gene expression; host organism interaction; inhibitor /antagonist; intermolecular interaction; method development; molecular genetics; phenotype; postdoctoral investigator; proteomics; small molecule; transposon /insertion element; virulence

DESCRIPTION (provided by applicant): The goal of this project is to further investigate the regulatory pathways that control virulence expression in Vibrio cholerae, a gram-negative bacterium that causes the human diarrheal disease called cholera. It also is a model organism for understanding bacterial pathogenesis, with the identification of many of its virulence factors and requirements for colonization. Yet, much remains unknown with regards to the organism's mechanisms to sense the environment and respond to virulence activating stimuli. Taking a chemical genetics approach to studying V. choIerae pathogenesis, the candidate has identified 11 novel small molecules that inhibit activation of the virulence genes in V. cholerae. Preliminary studies of these compounds suggest that they define regulatory mechanisms for virulence activation that have not been previously described, including mechanisms to respond to anaerobic conditions and to coordinate virulence expression with biofilm formation. The 11 compounds will be used to identify and characterize these novel pathways. In addition to genetic and biochemical approaches, two new methods will be developed for identifying regulators of virulence expression in V. cholerae. These two methods, a complementation method using a transposon based inducible promoter system and a bacterial-3-hybrid system for identifying small molecule-protein interactions, can be applied to the study of other bacterial systems. The long-term goal of the candidate is to merge the powerful fields of chemical and molecular genetics in the study of bacterial pathogenesis. In this era of increasing microbial resistance to current antibiotics, understanding the basic mechanisms of pathogenesis is critical to finding new solutions. Small molecule effectors and inhibitors may be developed as tools to probe the biology of microbes, providing insight into the organism's requirements for survival in the environment as well as for causing disease in the host. The short-term goal of the candidate is to acquire the skills of molecular bacterial genetics and postgenomic analysis in the laboratory of the sponsor, Dr. John Mekalanos, thus allowing her to become a productive, independent physician-scientist.

描述(申请人提供)：该项目的目标是进一步研究控制霍乱弧菌毒力表达的调控途径，霍乱弧菌是一种导致人类腹泻疾病霍乱的革兰氏阴性细菌。它也是了解细菌致病机理的模式生物，鉴定了它的许多毒力因素和定殖要求。然而，关于有机体感知环境和对毒力激活刺激做出反应的机制，仍有许多未知之处。 采用化学遗传学方法研究霍乱弧菌的致病机制，候选人已经确定了11个新的小分子，这些小分子可以抑制霍乱弧菌毒力基因的激活。对这些化合物的初步研究表明，它们定义了以前尚未描述的毒力激活调节机制，包括对厌氧条件做出反应的机制，以及协调毒力表达与生物膜形成的机制。 这11种化合物将被用来识别和表征这些新的途径。除了遗传和生化方法外，还将开发两种新的方法来鉴定霍乱弧菌毒力表达的调控因子。这两种方法，一种是基于转座子的可诱导启动子系统的互补法，另一种是鉴定小分子-蛋白质相互作用的细菌-3-杂交系统，这两种方法可以应用于其他细菌系统的研究。 候选人的长期目标是在细菌发病机制的研究中融合化学和分子遗传学的强大领域。在这个微生物对当前抗生素的耐药性日益增加的时代，了解发病的基本机制对于找到新的解决方案至关重要。小分子效应物和抑制物可被开发为探索微生物生物学的工具，从而洞察生物体在环境中生存的需求以及在宿主中引发疾病的需求。候选人的短期目标是在赞助商John Mekalanos博士的实验室获得分子细菌遗传学和后基因组分析的技能，从而使她成为一名多产的、独立的内科科学家。