Project 4. Defining cell division pathways in Mtb.

项目 4. 定义 Mtb 的细胞分裂途径。

• 批准号: 10426182
• 负责人: SABINE EHRT
• 金额: $ 70.46万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-12 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10426182
• 关键词: Animal Model; Bacillus subtilis; Bacteria; Biochemical; Cell Cycle; Cell Division Process; Cell Wall; Cell division; Cell model; Cell physiology; Cellular Metabolic Process; Cellular Structures; Collaborations; Complex; Core Facility; Cues; Cytokinesis; DNA biosynthesis; Data; Disease; Enzymes; Escherichia coli; Event; Genes; Genetic; Genetic Transcription; Genus Mycobacterium; Goals; Guanosine; Infection; Link; Mediator of activation protein; Metabolic; Metabolic Pathway; Metabolism; Methods; Morphogenesis; Mycobacterium tuberculosis; Nucleosides; Pathway Analysis; Pathway interactions; Phosphorylation; Process; Protein Kinase; Regulation; Role; Stress; Tuberculosis; Work; antimicrobial; base; cell assembly; cell growth; gene product; metabolomics; mycobacterial; periplasm; programs; protein complex; protein protein interaction; quantitative imaging; segregation

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.

项目 4，摘要 与所有细菌一样，结核分枝杆菌 (Mtb) 必须协调主要的细胞过程，例如 DNA 复制，通过结构成分的合成和分离来生长和分裂。虽然 控制细胞分裂的过程在模型生物体中得到了相当好的描述，其使用的机制 分枝杆菌明显不同，而且了解较少。定义管理基本要素的机制 结核分枝杆菌的细胞分裂过程将揭示新的抗菌靶点，并为理解这种作用如何提供基础 该过程在缓慢生长状态下受到调节，这有助于感染期间细菌的持续存在。这 该项目支持该计划的总体目标，即了解 Mtb 适应的重要途径 与疾病相关的压力。它将与其他项目协同并利用每个核心。 协调胞质分裂与细胞周期以及细胞物质的分布必须是暂时的 和空间调节。项目实验室的工作表明，这种协调至少需要三个不同的 监管范式。 (1) 细胞分裂复合物或“分裂体”的有序组装需要空间和 可能由 Ser/Thr 蛋白激酶 (STPK) 提供的时间线索。 (2)胞质外酶 负责细胞壁代谢的代表了一个独特的监管挑战，并且通常由蛋白质控制 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提供前体所必需的 这一重大细胞事件所需的。 该项目的目标是了解控制分枝杆菌细胞分裂的监管范式。 具体来说，该项目将： 目标 1. 表征蛋白质磷酸化在分裂动力学中的作用。同步山地车文化， 定量成像和生化方法将用于机械地表征 分裂功能的时间和空间调节中的磷酸化。 目标 2. 定义并表征细胞分裂所需的胞质外复合物。的组合 将使用遗传和物理方法来寻找对调节酶促重要的相互作用 细胞分裂过程中的活性和定位。 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需的主要代谢途径。