Regulation of VapC toxins by Ser/Thr phosphorylation of VapB antitoxins in M. tuberculosis

结核分枝杆菌中 VapB 抗毒素的 Ser/Thr 磷酸化对 VapC 毒素的调节

• 批准号: 9978276
• 负责人: ROBERT N HUSSON
• 金额: $ 26.55万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-21 至 2022-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9978276
• 关键词: Address; Affect; Affinity Chromatography; Bacteria; Bacterial Physiology; Bacterial Toxins; Binding; Biological Assay; Cell physiology; Cells; Cleaved cell; Communicable Diseases; DNA; Data; Development; Endoribonucleases; Family; Foundations; Gene Expression; Genes; Genus Mycobacterium; Goals; Granuloma; Growth; Infection; Investigation; Knowledge; Lead; Messenger RNA; Molecular; Morbidity - disease rate; Mycobacterium bovis; Mycobacterium tuberculosis; Operon; Pathogenesis; Pathway interactions; Phenotype; Phosphorylation; Phosphorylation Site; Phosphotransferases; Physiology; Play; Promoter Regions; Protein Kinase; Proteins; Proteome; RNA; Regulation; Research; Research Proposals; Ribosomal RNA; Role; Signal Transduction; Site; Stress; System; Technology; Testing; Toxin; Transfer RNA; Translations; Tuberculosis; Two-Hybrid System Techniques; Virulence; Work; antibiotic tolerance; antimicrobial tolerance; antitoxin; enzyme activity; human pathogen; insight; member; mortality; novel; novel strategies; promoter; receptor; response; stress tolerance; transcriptome sequencing

PROJECT SUMMARY The long-term goal of this research is to understand how Mycobacterium tuberculosis toxin-antitoxin systems are regulated to contribute to tuberculosis pathogenesis. The specific goal of this R21 research proposal is to investigate our hypothesis that Ser/Thr phosphorylation of VapB antitoxins by receptor-type Ser/Thr protein kinases represents a novel mechanism by which VapC toxins can be regulated in response to environmental signals. VapC toxins are the majority of all TA toxins in M. tuberculosis and have been implicated in proteome remodeling, growth arrest, antibiotic tolerance and ability to survive a range of stresses that are relevant for tuberculosis pathogenesis. The established role of VapB antitoxins in sequestering cognate VapC toxin proteins and autoregulating vapBC gene expression, together with our recent finding of significantly decreased phosphorylation of several VapB proteins in the setting of specific kinase depletion, provide the premises for this research. This research proposal has two Aims. In Aim 1 we will determine the effects of phosphorylation of two VapB antitoxins on i) the growth of M. tuberculosis expressing these antitoxins together with their cognate VapC toxins, and ii) the molecular interactions of these VapB toxins with their cognate VapC proteins and their cognate promoters. In Aim 2 we will investigate the effects of phosphorylation of these VapB antitoxins on VapC enzyme activity. This research will test our hypothesis that VapB Ser/Thr phosphorylation may be a means to regulate VapC activity and will begin to characterize the molecular mechanism(s) by which this regulation occurs. Results from this research will provide the foundation for further investigation into how Ser/Thr phosphorylation of VapB antitoxins can function to transduce signals to control the activity of VapC toxins that are relevant for M. tuberculosis growth control, stress tolerance and tuberculosis pathogenesis.

项目摘要 这项研究的长期目标是了解结核分枝杆菌毒素 - 抗毒素如何 系统受调节为有助于结核病发病机理。这项R21研究的具体目标 提案是研究我们的假设，即受体类型对VAPB抗毒素的Ser/Thr磷酸化 Ser/Thr蛋白激酶代表了一种新的机制，可以通过该机制来调节VAPC毒素 环境信号。 VAPC毒素是结核分枝杆菌中所有TA毒素中的大多数 与蛋白质组的重塑，生长停滞，抗生素耐受性和生存能力有关 与结核病发病机理有关。 VAPB抗毒素在隔离中的既定作用 同源VAPC毒素蛋白和自动调节VAPBC基因表达，以及我们最近的发现 在特定激酶消耗的情况下，几种VAPB蛋白的磷酸化显着降低， 提供本研究的前提。 该研究建议有两个目标。在AIM 1中，我们将确定两个磷酸化的影响 i）抗毒素的抗毒素i）结核分枝杆菌的生长表达这些抗毒素及其同源 VAPC毒素，以及II）这些VAPB毒素与它们的同源VAPC蛋白及其的分子相互作用 同源启动子。在AIM 2中，我们将研究这些VAPB抗毒素对磷酸化的影响 VAPC酶活性。 这项研究将检验我们的假设，即VAPB Ser/Thr磷酸化可能是调节的一种手段 VAPC活性并将开始表征该调节的分子机制。 这项研究的结果将为进一步研究Ser/Thr磷酸化的基础提供基础 VAPB抗毒素的作用可以转导信号以控制与与 结核病生长控制，胁迫和结核病发病机理。