Dissection of Mycobacterium tuberculosis metabolic and regulatory pathways to per

描述（由申请人提供）：该计划的目标是确定与M相关的关键代谢途径/反应和调节转录因子。结核生长停滞（与持续感染有关的生理状态）。通过转录谱分析对M.在持续性感染的鼠模型中的结核病碳代谢导致了一个假设，即在持续性状态的建立期间发生协调的代谢转化。它代表了生长和生长停滞杆菌中截然不同的碳流模式。虽然碳用于能量产生和生物合成前体用于生长细胞中的细菌生长;但在持久性细胞中，碳主要用于形成储存化合物。为了验证这一假设，13 C同位素标记实验结合代谢网络建模将用于通过测量同位素异构体分布到其前体代谢物中的生长和非生长稳态培养物来确定各种途径/反应之间的碳流。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。第二，利用报告基因融合和转座子突变技术研究关键代谢基因是否以协同方式调控。这种方法将导致识别的转录因子，调节关键代谢基因在持久性。该项目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：该计划的目标是确定在建立M。结核持续感染。该项目有望为结核杆菌的持续感染机制提供新的认识，并为解决持续感染提供新的策略。

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.