PodJ processing and polar development in Caulobacter crescentus

新月柄杆菌的 PodJ 加工和极性发育

• 批准号: 7600533
• 负责人: Patrick David Curtis
• 金额: $ 4.72万
• 依托单位: TRUSTEES OF INDIANA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7600533
• 关键词: Affect; Animals; Bacteria; Bacterial Physiology; Behavior; Binding; Biochemical; Biochemistry; Biological Assay; Biological Models; Biology; Caulobacter crescentus; Cell Cycle; Cell Polarity; Cell division; Cells; Cellular biology; Chemotaxis; Cytoskeleton; Data; Development; Developmental Process; Disease; Elements; Event; Genetic; Genetic Screening; Goals; Knowledge; Lead; Length; Location; Mastigophora; Mediating; Methods; Mutagenesis; Mutation; Organelles; Organism; Pathogenicity; Peptidoglycan; Phenotype; Pilum; Plants; Polymers; Process; Prokaryotic Cells; Proteins; Proteolytic Processing; Research; Role; Site; Site-Directed Mutagenesis; Solid; Sorting - Cell Movement; Staging; Structure; System; Techniques; Virulence; Virulence Factors; Work; cell motility; combat; mutant; pathogen; periplasm; research study; sortase

DESCRIPTION (provided by applicant): The long-term goal of this research is to determine how bacterial cells establish intracellular polarity, direct specific processes to the poles, and control the activity of polar processes. Many bacteria target organelles and/or processes specifically to the cell poles. Particularly, it has been shown that many pathogens polarly localize virulence factors, indicating that polar localization has implications in disease. The basic mechanisms for polar localization are unknown. In this work, development of Caulobacter crescentus is used as a model system for polarity. Polar development in this organism has been correlated with the subcellular localization and proteolytic processing of the protein PodJ, and C. crescentus cells that are inhibited in development show no PodJ processing, indicating that the stability of PodJ is tied to polar developmental processes. Other proteins similar to PodJ have been found in other organisms, including pathogens that demonstrate cellular polarity. By understanding the role of PodJ in controlling polar processes, it may be possible to extrapolate this knowledge to other bacterial systems. The specific aims of this work are: 1. generate mutations in PodJ that lead to increased protein stability and determine the affect of increased stability of PodJ-mediated polar processes, and 2. determine the role of the putative PodJ peptidoglycan binding domain in maintaining PodJ stability. A combination of site-directed mutagenesis of sequences predicted to impact PodJ stability, and unbiased genetic screens using powerful assays to identify PodJ stability mutants will be utilized. Analysis of mutant phenotypes will involve cell biology techniques, biochemistry techniques, and bacterial physiology assays. Several plant and animal pathogens demonstrate cellular polarity and polar localization of virulence factors, yet the basic mechanisms of bacterial polarity are unknown. Understanding the underlying methods of establishing and controlling bacterial polarity will facilitate easier access to specific strategies employed by pathogens, and enhance new strategies to combat their virulence.

描述（由申请人提供）：这项研究的长期目标是确定细菌细胞如何建立细胞内极性，直接到杆的特定过程并控制极性过程的活性。许多细菌靶向细胞器和/或过程，专门针对细胞杆。特别是，已经表明，许多病原体两极分化的毒力因子，表明极地定位对疾病有影响。极性定位的基本机制尚不清楚。在这项工作中，花椰菜齿轮的发展用作极性模型系统。该生物体的极性发育与蛋白质PODJ的亚细胞定位和蛋白水解处理相关，并且在发育中抑制的新月形细胞没有显示PODJ加工，表明PODJ的稳定性与极性发育过程有关。在其他生物体中发现了类似于PODJ的其他蛋白质，包括证明细胞极性的病原体。通过了解PODJ在控制极性过程中的作用，可以将这些知识推断到其他细菌系统。这项工作的具体目的是：1。在PODJ中产生突变，从而导致蛋白质稳定性提高，并确定PODJ介导的极性过程的稳定性提高的影响，以及2。确定推定的PODJ podjpeptidoglycan结合结构域在维持PODJ稳定性方面的作用。将利用使用强大的测定法来识别PODJ稳定性突变体的序列的位置定向诱变，以及无偏的遗传筛选。突变表型的分析将涉及细胞生物学技术，生物化学技术和细菌生理学测定。几种动植物病原体表现出毒力因子的细胞极性和极性定位，但细菌极性的基本机制尚不清楚。了解建立和控制细菌极性的基本方法将有助于更容易获得病原体采用的特定策略，并增强对抗其毒力的新策略。