Dissection of Mycobacterium tuberculosis metabolic and regulatory pathways to per

描述（由申请人提供）：该项目的目标是确定与结核分枝杆菌生长停滞（一种与持续感染相关的生理状态）相关的关键代谢途径/反应和调节转录因子。通过对小鼠持续感染模型中结核分枝杆菌碳代谢转录谱的初步研究，得出了在持续状态建立过程中发生协调代谢转化的假设。它代表了生长和生长停滞杆菌中截然不同的碳流模式。而碳则被用于能量产生和生物合成前体，供细菌在生长的细胞中生长；在持久性细胞中，碳主要被动员起来形成储存化合物。为了验证这一假设，13C同位素标记实验将结合代谢网络模型，通过测量生长和非生长稳态培养中同位素体到其前体代谢物的分布，来确定各种途径/反应中的碳流动。这种方法将允许识别与细菌持久性相关的关键代谢途径/反应。其次，利用报告基因融合和转座子诱变技术，研究关键代谢基因是否以协同模式调控。这种方法将导致鉴定在持续过程中调节关键代谢基因的转录因子。该方案有望为结核杆菌的持续感染机制提供新的见解，并为解决持续感染提供可能的策略。公共卫生相关性：该项目的目标是确定结核分枝杆菌持续性感染建立过程中的关键代谢和调节事件。该计划预计将导致新的知识关于结核杆菌的持续机制，并提供新的策略，以解决持续感染。

DESCRIPTION (provided by applicant): The goal of the program is to identify the key metabolic pathways/reactions and regulatory transcription factors that are associated with M. tuberculosis growth arrest (a physiological state related to persistent infection). Preliminary study by transcription profiling of M. tuberculosis carbon metabolism in a murine model of persistent infection led to a hypothesis that a coordinated metabolic transformation occurs during the establishment of persistent state. It represents a drastic different carbon flow patterns in growing and growth-arrested bacilli. While carbons are utilized for energy generation and biosynthetic precursors for bacterial growth in growing cells; in persistent cells carbons are mainly mobilized for the formation of storage compounds. To test the hypothesis, 13C isotope-labeling experiment in combination with metabolic network modeling will be used to determine carbon flows among various pathways/reactions by measuring isotopomer distribution to their precursor metabolites in growing and nongrowing steady-state cultures. This approach will allow the identification key metabolic pathways/reactions associated with bacterial persistence. Second, reporter fusion and transposon mutagenesis technology will be utilized to study whether key metabolic genes are regulated in a coordinated mode. This approach will lead to identification of transcription factors that regulate key metabolic genes during persistence. This program is expected to provide new insights into the persistent mechanism of tubercle bacilli and offer possible strategies to tackle the persistent infection. PUBLIC HEALTH RELEVANCE: The goal of the program is to identify the key metabolic and regulatory events during the establishment of M. tuberculosis persistent infection. The program is expected to lead to new knowledge about the persistent mechanism of the tubercle bacilli and offer new strategies to tackle the persistent infection.