The regulation of c-di-AMP homeostasis and signaling in bacteria

细菌中 c-di-AMP 稳态和信号传导的调节

• 批准号: 10707941
• 负责人: Tu Anh Huynh
• 金额: $ 38.88万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-21 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10707941
• 关键词: Anti-Bacterial Agents; Bacteria; Biochemical; Biological Assay; Biological Process; Cells; Defect; Detection; Development; Firmicutes; Genetic; Genetic Engineering; Growth; Homeostasis; Hydrolysis; Laboratories; Laboratory Finding; Molecular Target; Nucleotides; Pathway interactions; Phenotype; Physiology; Regulation; Role; Second Messenger Systems; Signal Transduction; Toxic effect; biological adaptation to stress; fitness; mutant; novel; phosphoric diester hydrolase; small molecule

PROJECT SUMMARY The small nucleotide c-di-AMP is a ubiquitous second messenger produced by thousands of bacterial and archeal species. Many bacteria, especially those in the Firmicutes phylum, require c-di-AMP for growth in standard laboratory media, and the mechanisms for c-di-AMP essentiality have been extensively studied. However, in a wide range of c-di-AMP-producing bacteria, including those for which c-di-AMP is completely dispensable, unregulated c-di-AMP accumulation is toxic, diminishing bacterial growth and stress response. Thus, c-di-AMP homeostasis is critical for bacteria that produce it. Nevertheless, the mechanisms that regulate c-di-AMP levels in the bacterial cell, and the mechanisms by which c-di-AMP regulates different cellular pathways to achieve optimal physiology, are both poorly understood. My lab found that c-di-AMP levels are dynamic in the bacterial cell, and c-di-AMP hydrolysis is a key mechanism to modulate c-di-AMP levels. Furthermore, we have generated bacterial mutants lacking c-di-AMP phosphodiesterases, called pde mutants, and extensively studied their phenotypic defects. In the next five years, we will investigate two major aspects of c-di-AMP signaling: i) the signal detection and regulatory mechanisms of c-di-AMP phosphodiesterase, and ii) the mechanisms by which c-di-AMP coordinates its molecular targets to achieve optimal physiology and stress response. For the second theme, we will take two complementary approaches. One, we will elucidate the biological functions of c- di-AMP molecular targets based on targeted phenotypic, genetic, and biochemical analyses. Two, we will identify the molecular targets underlying the toxicity phenotypes of pde mutants, through genetic fitness assays. Our findings will reveal fundamental mechanisms of c-di-AMP signaling, and these mechanisms will provide a framework to understand the global regulatory roles of c-di-AMP in the physiology of different bacterial species.

项目摘要 小核苷酸c-di-AMP是由数千种细菌和微生物产生的普遍存在的第二信使， 原始物种许多细菌，特别是厚壁菌门中的那些细菌，需要c-di-AMP以在细胞中生长。 标准实验室培养基，和c-di-AMP重要性的机制已被广泛研究。 然而，在广泛的产生c-di-AMP的细菌中，包括那些c-di-AMP完全被 然而，不受调节的c-di-AMP积累是有毒的，减少细菌生长和应激反应。 因此，c-di-AMP稳态对于产生它的细菌是至关重要的。 细菌细胞中的c-di-AMP水平，以及c-di-AMP调节不同细胞途径的机制 以达到最佳的生理状态，都知之甚少。我的实验室发现c-di-AMP的水平是动态的 细菌细胞中，c-di-AMP水解是调节c-di-AMP水平的关键机制。此外，我们还 产生了缺乏c-di-AMP磷酸二酯酶的细菌突变体，称为pde突变体，并进行了广泛的研究 他们的表型缺陷在接下来的五年里，我们将研究c-di-AMP信号传导的两个主要方面： c-di-AMP磷酸二酯酶的信号检测和调节机制，以及ii）通过 其中c-di-AMP协调其分子靶点以实现最佳生理和应激反应。为 第二个主题，我们将采取两种相辅相成的办法。首先，我们将阐明c的生物学功能- 基于靶向表型、遗传和生物化学分析的di-AMP分子靶标。第二，我们将确定 通过遗传适应性测定，研究PDE突变体毒性表型的分子靶点。我们 研究结果将揭示c-di-AMP信号传导的基本机制，这些机制将提供一个 该框架旨在了解c-di-AMP在不同细菌物种生理学中的全局调节作用。