Project 4. Defining cell division pathways in Mtb.

项目4，摘要 像所有细菌一样，结核分枝杆菌（Mtb）必须协调主要的细胞过程，如DNA 复制，通过合成和分离结构成分以生长和分裂。而 控制细胞分裂的过程在模式生物中得到了合理的描述， 分枝杆菌是明显不同的，并且不太清楚。界定管理基本人权的机制 Mtb的细胞分裂过程将揭示新的抗菌靶标，并为理解这种作用机制提供基础。 这一过程在缓慢生长状态期间受到调节，这有助于感染期间细菌的持久性。这 该项目支持该计划的总体目标，以了解结核病适应的重要途径， 疾病相关的压力。它将与其他项目协同并利用每个核心。 细胞质分裂与细胞周期的协调以及细胞物质的分布必须是暂时的， 并且空间上受到调节。项目实验室的工作表明，这种协调至少需要三个不同的 监管模式。(1)细胞分裂复合体或“分裂体”的有序组装需要空间和生物学上的相互作用。 可能由丝氨酸/苏氨酸蛋白激酶（STPK）提供的时间线索。(2)细胞质外酶 对细胞壁代谢负责是一个独特的调节挑战， 蛋白质在周质空间的相互作用。(3)细胞分裂所必需的过程的协调是 不限于隔膜;基本的代谢变化也可能需要提供前体 这一重大细胞事件所必需的。 这个项目的目标是了解控制分枝杆菌细胞分裂的调控模式。 具体而言，该项目将： 目标1。描述蛋白质磷酸化在分裂体动力学中的作用。同步结核菌培养， 定量成像和生物化学方法将用于机械表征的作用， 磷酸化在分裂体功能的时间和空间调节中的作用。 目标2.定义和表征细胞分裂所需的胞质外复合物。的组合 遗传和物理方法将用于发现对调节酶的重要的相互作用。 在细胞分裂期间的活性和定位。 目标3。描述细胞分裂和新陈代谢之间的联系。代谢组学和 遗传学将用于研究细胞分裂所必需的主要代谢途径。

Project 4, Abstract Like all bacteria, Mycobacterium tuberculosis (Mtb) must coordinate major cellular processes, such as DNA replication, with the synthesis and segregation of structural components in order to grow and divide. While the processes governing cell division are reasonably well-described in model organisms, the mechanisms used by mycobacteria are clearly different and less well understood. Defining the mechanisms governing the essential process of cell division in Mtb will reveal new antimicrobial targets and provide the basis to understand how this process is regulated during the slow-growing states that contribute to bacterial persistence during infection. This project supports the program’s overall goal to understand pathways important to Mtb’s adaptation to disease-relevant stress. It will synergize with other projects and leverage each of the cores. Coordinating the cytokinesis with the cell cycle and the distribution of cellular material must be both temporally and spatially regulated. Work in the project labs has shown that this coordination requires at least three distinct regulatory paradigms. (1) The ordered assembly of the cell division complex, or “divisome” requires spatial and temporal cues that may be provided by Ser/Thr protein kinases (STPK). (2) Extracytoplasmic enzymes responsible for cell wall metabolism represent a distinct regulatory challenge, and are often controlled by protein- protein interactions in the periplasmic space. (3) The coordination of processes necessary for cell division are not restricted to the septum; fundamental metabolic changes are also likely necessary to provide the precursors required for this major cellular event. The goal of this project is to understand the regulatory paradigms that control mycobacterial cell division. Specifically, the project will: Aim 1. Characterize the role of protein phosphorylation in divisome dynamics. Synchronized Mtb cultures, quantitative imaging, and biochemical approaches will be used to mechanistically characterize the role of phosphorylation in the temporal and spatial regulation of divisome function. Aim 2. Define and characterize extracytoplasmic complexes necessary for cell division. A combination of genetic and physical approaches will be used to find interactions that are important for regulating enzymatic activity and localization during cell division. Aim 3. Characterize the links between cell division and metabolism. A combination of metabolomics and genetics will be used to investigate the primary metabolic pathways necessary for cell division.