The role of cyclic-di-AMP in the regulation of Chlamydia cellular functions

环二AMP在衣原体细胞功能调节中的作用

• 批准号: 8776266
• 负责人: Raphael H Valdivia
• 金额: $ 19.63万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-12-01 至 2016-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8776266
• 关键词: 5' Untranslated Regions; Acylation; Address; Affinity Chromatography; Applications Grants; Binding; Binding Proteins; Biochemical; Biochemical Genetics; Biological Assay; Biology; Cell Wall; Cell membrane; Cell physiology; Chlamydia; Chlamydia trachomatis; Collection; Cues; Development; Diffusion; Dinucleoside Phosphates; Enzyme Gene; Escherichia coli; Gene Expression; Genes; Genetic; Gram-Negative Bacteria; Gram-Positive Bacteria; Health; Homeostasis; Integral Membrane Protein; Laboratories; Libraries; Life Cycle Stages; Mass Spectrum Analysis; Mediating; Messenger RNA; Methods; Microbial Biofilms; Molecular; Monitor; Nucleic Acids; Operon; Pathogenesis; Proteins; Proteomics; RNA; Radiolabeled; Recombinant Proteins; Regulation; Reporter Genes; Research; Role; Sexually Transmitted Diseases; Signal Transduction; Testing; Trachoma; Virulence; Work; base; cellular development; clinically relevant; genetic analysis; genetic approach; genome-wide analysis; microbial; mutant; pathogen; programs; radiotracer; research study; response; reverse genetics

DESCRIPTION (provided by applicant): Chlamydia trachomatis, the causative agent of blinding trachoma and many sexually transmitted diseases, exists in two developmental forms: an infectious elementary body (EB) and a replicative reticulate body (RB). The transition between EB and RB forms is an essential step in the life cycle of this obligate bacterial pathogen. However, remarkably little is known about the environmental cues and molecular mechanism underlying these developmental changes. We recently determined that C. trachomatis EBs accumulate cyclic di-adenosyl monophosphate (c-di-AMP), a newly identified nucleic acid metabolite that contributes to cell wall homeostasis in gram-positive bacteria. In gram-negative bacteria, the structurally related cyclic dinucleotide c-di-GMP is an allosteric regulator of enzymes, gene expression and riboswitches important for biofilm formation and microbial virulence. We hypothesize that c-di-AMP regulates protein and/or RNA functions to promote the RB to EB transition and possibly virulence. In this exploratory research program, we outline experiments to identify proteins that regulate the Chlamydia diadenylate cyclase, and protein(s) and RNA(s) that may be targets of c-di-AMP-mediated regulation. We will employ new biochemical and genetic methods developed in our laboratory to assess the role of c-di-AMP and its identified target(s) in chlamydial development and pathogenesis.

描述（由申请人提供）：沙眼衣原体是致盲性沙眼和许多性传播疾病的病原体，以两种发育形式存在：传染性基本体（EB）和复制性网状体（RB）。 EB 和 RB 形式之间的转变是这种专性细菌病原体生命周期中的重要步骤。然而，人们对这些发育变化背后的环境因素和分子机制知之甚少。我们最近确定沙眼衣原体 EB 会积累环状二腺苷单磷酸 (c-di-AMP)，这是一种新发现的核酸代谢物，有助于革兰氏阳性菌的细胞壁稳态。在革兰氏阴性细菌中，结构相关的环状二核苷酸 c-di-GMP 是酶、基因表达和核糖开关的变构调节剂，对生物膜形成和微生物毒力至关重要。我们假设 c-di-AMP 调节蛋白质和/或 RNA 功能，以促进 RB 向 EB 的转变，并可能促进毒力。在这个探索性研究计划中，我们概述了鉴定调节衣原体二腺苷酸环化酶的蛋白质以及可能是 c-di-AMP 介导的调节目标的蛋白质和 RNA 的实验。我们将采用我们实验室开发的新生化和遗传学方法来评估 c-di-AMP 及其确定的靶标在衣原体发育和发病机制中的作用。