Molecular Genetic Analysis of Chlamydia Pathogenicity

衣原体致病性的分子遗传学分析

• 批准号: 8707934
• 负责人: Anthony T Maurelli
• 金额: $ 37.81万
• 依托单位: HENRY M. JACKSON FDN FOR THE ADV MIL/MED
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-12-01 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8707934
• 关键词: Acute; Address; Alanine Racemase; Amino Acids; Animals; Bacteria; Biochemical; Biochemical Genetics; Biology; Biotin; Blindness; Cell Wall; Cells; Cervicitis; Chlamydia; Chlamydia Infections; Chlamydophila psittaci; Cloning; Complex; Cytosol; Development; Disease; Endocarditis; Enzyme Gene; Enzymes; Epididymitis; Female; Funding; Genes; Genetic; Genital system; Glutamate racemase; Goals; Growth; Immune response; Infertility; Iron; Knowledge; Laboratories; Lead; Learning; Libraries; Life Style; Light; Link; Lung; Metabolic Pathway; Metabolism; Molecular; Molecular Genetics; Nonspecific urethritis; Nutrient; Organism; Pathogenesis; Pathogenicity; Pathway interactions; Pelvic Inflammatory Disease; Peptidoglycan; Physiology; Plasmids; Pneumonia; Polyarthritides; Prevention; Process; Reproductive Health; Research; Role; Salpingitis; Sexually Transmitted Diseases; Shigella; Shuttle Vectors; Signal Transduction; Structure; System; Techniques; Testing; Therapeutic Intervention; Transgenes; Vitamins; Woman; Work; antimicrobial; genetic analysis; innovation; insight; male; man; mutant; new technology; pathogen; public health relevance; success; technology development; tool; tool development; uptake

DESCRIPTION (provided by applicant): Bacteria of the genus Chlamydia are significant pathogens of animals and man. The diseases caused by Chlamydia spp. in man include pneumonia, endocarditis, polyarthritis, blindness, and a wide range of sexually transmitted diseases including cervicitis, salpingitis, pelvic inflammatory disease, and infertility in females and non-gonococcal urethritis and acute epididymitis in males. Despite many years of effort, the Chlamydia remain intractable to genetic analysis due to their obligate intracellular lifestyle and complex developmental cycle. Our long-term goal is to apply the full range of molecular tools including the power of genetics to study the pathogenic mechanisms and intracellular metabolism of Chlamydia. The aims of this proposal include a technology development aim (genetic tools) and two hypothesis-driven aims to address significant questions of Chlamydia biology. The specific aims are: 1 - Development of tools for the genetic analysis of Chlamydia. 2 - Biochemical and genetic characterization of peptidoglycan synthesis in Chlamydia. 3 - Identification of transport systems for uptake of essential constituents for Chlamydia growth. We will employ new and innovative approaches to build on our past success and develop genetic tools in aim 1. Success in achieving this aim will have a significant impact and advance the field of Chlamydia research by making new tools available for genetic analysis of Chlamydia. Aim 2 will identify the enzymes involved in two key cytoplasmic steps in peptidoglycan (PG) synthesis and also apply new techniques to isolate and demonstrate the presence of PG components in Chlamydia. We will also screen for Chlamydia mutants defective in signaling to the host via PG fragments as a first step in dissecting the role of PG in host response. Success in aim 2 will finally resolve the "Chlamydia anomaly" and reveal potential new targets for antimicrobial development. Aim 3 will characterize transport systems used by Chlamydia to acquire iron and biotin from the host cytosol. Mechanisms of iron acquisition by Chlamydia are completely unknown and our innovative approach will use a Shigella mutant to screen a Chlamydia library. Since the genes for biotin synthesis are absent in C. trachomatis, transport of this essential vitamin is critical for growth. In both cases, success in this aim will shed light on processes of nutrient acquisition that are potential targets for therapeutic intervention. In broader terms, the significance of this aim is that it will provide insight into metabolic processes of obligate intracellular pathogens.

描述（由申请人提供）：衣原体属的细菌是动物和人的重要病原体。衣原体属引起的疾病。人类包括肺炎，心内膜炎，多重关节炎，失明以及广泛的性传播疾病，包括宫颈炎，分肠疾病，骨盆炎性疾病以及女性的不育症，男性非造成肌肉炎和非造成病毒性尿道炎和急性表育体。尽管经过多年的努力，衣原体由于其强大的细胞内生活方式和复杂的发育周期而对遗传分析仍然很棘手。我们的长期目标是应用各种分子工具，包括遗传学的力量来研究衣原体的致病机制和细胞内代谢。该提案的目的包括技术开发目标（遗传工具）和两个以假设为导向的旨在解决衣原体生物学的重要问题。具体目的是：1-开发衣原体遗传分析的工具。 2-衣原体中肽聚糖合成的生化和遗传表征。 3-鉴定运输系统以吸收衣原体生长的基本成分。我们将采用新的和创新的方法来建立我们过去的成功并在AIM 1中开发遗传工具。实现这一目标的成功将对衣原体研究产生重大影响，并通过为衣原体的遗传分析提供新的工具。 AIM 2将鉴定肽聚糖（PG）合成两个关键细胞质步骤中涉及的酶，并应用新技术隔离并证明在衣原体中存在PG成分。我们还将通过PG片段向宿主发出信号传导时缺陷的衣原体突变体，作为解剖PG在宿主反应中的作用的第一步。 AIM 2的成功最终将解决“衣原体异常”，并揭示潜在的抗菌发育目标。 AIM 3将表征衣原体用来从宿主细胞质中获取铁和生物素的运输系统。衣原体采购铁的机制是完全未知的，我们的创新方法将使用志贺氏菌突变体来筛选衣原体图书馆。由于沙眼梭状芽孢杆菌不存在生物素合成的基因，因此这种必需维生素的转运对于生长至关重要。在这两种情况下，这一目标的成功都将揭示营养习得的过程，这是治疗干预的潜在目标。从更广泛的角度来看 该目标的重要性是，它将洞悉对强制性细胞内病原体的代谢过程。