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，与结构成分的合成和分离，以生长和分裂。而当 管理细胞分裂的过程在模型生物中得到了相当好的描述， 分枝杆菌显然是不同的，也不太为人所知。定义管理本质的机制 结核分枝杆菌的细胞分裂过程将揭示新的抗微生物靶点，并为理解这一过程提供基础 过程是在缓慢生长的状态下调节的，这有助于细菌在感染期间的持久性。这 该项目支持该计划的总体目标，即了解对结核分枝杆菌适应很重要的途径 与疾病相关的压力。它将与其他项目协同，并利用每个核心。 胞质分裂与细胞周期和细胞物质分布的协调必须是暂时的 并在空间上受到控制。项目实验室的工作表明，这种协调至少需要三个不同的 监管范例。(1)细胞分裂复合体或“分裂体”的有序组装需要空间和 可能由丝氨酸/苏氨酸蛋白激酶(STPK)提供的时间线索。(2)胞质外酶 负责细胞壁新陈代谢是一个明显的调节挑战，通常由蛋白质控制- 周质空间中的蛋白质相互作用。(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是必要的，以提供前体 这是这一重大细胞活动所必需的。 这个项目的目标是了解控制分枝杆菌细胞分裂的调控范例。 具体而言，该项目将： 目的1.研究蛋白质磷酸化在分裂组动力学中的作用。同步的MTB文化， 定量成像和生化方法将被用来机械地表征 在分裂体功能的时空调节中的磷酸化。 目的2.定义和表征细胞分裂所必需的胞外复合体。一种组合 基因和物理方法将被用来寻找对酶的调节很重要的相互作用 细胞分裂过程中的活性和定位。 目的3.描述细胞分裂和新陈代谢之间的联系。代谢组学和新技术的结合 遗传学将被用来研究细胞分裂所需的主要代谢途径。

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.