Peptidoglycan Assembly, Degradation, and Function in Pathogenic Chlamydia

致病性衣原体中肽聚糖的组装、降解和功能

基本信息

批准号：
10062849
负责人：
Anthony T Maurelli
金额：
$ 38.13万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-12-05 至 2022-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10062849
关键词：
Alleles Animals Antibiotics Architecture Area Bacteria Bacterial Infections Bacterial Sexually Transmitted Diseases Biochemical Biological Assay Biology Blindness Cell Shape Cell Wall Cell division Cells Cellular Morphology Chlamydia Chlamydia Infections Chlamydia genome Chlamydia trachomatis Complement Conflict (Psychology)Development Disease Enzymes Fluorescence Microscopy Gene Expression Genes Genetic Genetic Transformation Genital Genitalia Goals Gram-Negative Bacteria Growth Human Immune Immune response Immune system Inflammation Inflammatory Response Innate Immune Response Innate Immune System Knock-out Knowledge Label Life Cycle Stages Lung Maintenance Mass Spectrum Analysis Metabolic Methods Microbe Microscopy Modification Molecular Mutagenesis Osmotic Pressure Pathogenicity Pathologic Processes Pathway interactions Pelvic Inflammatory Disease Peptidoglycan Phase Physiology Play Polysaccharides Population Predisposition Prevention Process Production Recycling Regulation Reporter Reporting Reproductive Health Research Research Personnel Resolution Role Signaling Molecule Stress Structure Suggestion Techniques Technology Testing Thinness Tissues Translating Woman antimicrobial enzyme biosynthesis extracellular fluorescence imaging host-microbe interactions insight member novel therapeutics pathogenic bacteria periplasm polymerization promoter protein complex public health relevance receptor response trait vector

项目摘要

ABSTRACT Chlamydiae are obligate intracellular bacterial pathogens that cause disease in human and animal populations worldwide. C. trachomatis is the most prominent cause of both bacterial sexually transmitted disease and infectious blindness in the world. Peptidoglycan (PG) plays a critical role in the physiology of all bacteria. It determines cell morphology, provides protection against osmotic stress, and plays a critical role in cell division. Fragments of PG are also recognized by mammalian receptors and stimulate the host inflammatory response during bacterial infection. Although recent metabolic labeling techniques finally succeeded in demonstrating the presence of PG in Chlamydia, large gaps in our knowledge still remain. What role does PG play in cell division? How are PG glycan chains assembled in the chlamydial periplasm? How does Chlamydia limit the production of degradation fragments that can stimulate the innate immune response? Chlamydia infection in both the ocular as well as genital niches induces a severe inflammatory response that leads to tissue damage including blindness and pelvic inflammatory disease. It is unclear to what extent components of Chlamydia PG are responsible for this response. We need more insight into how Chlamydia synthesizes and degrades its peptidoglycan to better understand the pathologic processes of Chlamydia disease. The long-term goal is to determine the functions of peptidoglycan in Chlamydia physiology, specifically in cell division and development. The central hypothesis to be tested is that the PG synthesized by Chlamydia plays critical roles in cell division and the host immune response. A major focus will be on PG assembly, degradation, and recycling, which we hypothesize are central to these processes. The interrelated Specific Aims of this proposal are: 1. Elucidate the role played by PG in chlamydial cell division 2. Identify and characterize the genes involved in assembly/polymerization and degradation of chlamydial PG 3. Determine the fate and immunostimulatory potential of chlamydial PG muropeptides subsequent to their degradation Breakthrough technologies in experimental manipulation of Chlamydia make our proposal feasible: genetic transformation of Chlamydia, complementation vectors, inducible promoter constructs for controlled gene expression in Chlamydia, and allelic exchange/knockout mutagenesis. We will also employ metabolic labeling of PG, immune-reporter assays, mass spectroscopy, and superresolution microscopy. This research will fill in critical gaps in our understanding of Chlamydia growth and its recognition by host cells. The knowledge gained will provide information that may translate into new drugs to inhibit chlamydial PG synthesis or disrupt the PG fragments that can trigger the severe inflammation that accompanies Chlamydia infection.

抽象的衣原体是务本细胞细菌病原体，引起人类和动物的疾病全球人口。沙眼梭状芽胞庭是两种细菌性传播的最突出的原因世界上的疾病和感染性失明。肽聚糖（PG）在所有人的生理学中起着至关重要的作用细菌。它决定了细胞形态，提供了防止渗透压力的保护，并在细胞分裂。 PG的片段也被哺乳动物受体识别并刺激宿主细菌感染期间的炎症反应。虽然最近的代谢标签技术终于成功地证明了PG在衣原体中的存在，我们的知识仍然存在很大的差距。什么 PG在细胞部门中扮演角色吗？ PG聚糖链如何在衣原体的周围聚集？如何衣原体是否限制了可以刺激先天免疫反应的降解片段的产生？眼部和生殖器壁ches中的衣原体感染都会引起严重的炎症反应导致组织损伤，包括失明和骨盆炎症性疾病。目前尚不清楚多大程度上衣原体PG的组成部分负责此反应。我们需要更多地了解衣原体综合并降解其肽聚糖，以更好地了解衣原体疾病。长期目标是确定肽聚糖在衣原体生理学中的功能，特别是在细胞分裂和开发中。要测试的中心假设是由衣原体在细胞分裂和宿主免疫反应中起关键作用。重点将放在PG上组装，降解和回收利用，我们认为这对于这些过程至关重要。该提议相互关联的具体目的是： 1。阐明PG在衣原体细胞部门中所扮演的角色 2。识别和表征与衣原体PG组装/聚合和降解有关的基因 3。确定衣原体PG杂肽的命运和免疫刺激潜力降解衣原体实验操纵中的突破性技术使我们的建议可行：衣原体的遗传转化，互补载体，可诱导的启动子构建体衣原体中的基因表达以及等位基因交换/敲除诱变。我们还将使用代谢 PG的标记，免疫培养基测定法，质谱和超分辨率显微镜的标记。这项研究将填补我们对衣原体生长及其宿主细胞认识的关键空白。这获得的知识将提供可能转化为新药以抑制衣原体PG合成的信息或破坏可能触发衣原体感染的严重炎症的PG碎片。