Regulation of RecA Intein splicing in M. tuberculosis

结核分枝杆菌中 RecA 内含肽剪接的调控

基本信息

批准号：
8598326
负责人：
Kathleen A McDonough
金额：
$ 20万
依托单位：
WADSWORTH CENTER
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-05-25 至 2015-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8598326
关键词：
Address Affect Antineoplastic Agents Antitubercular Agents Bacteria Behavior Biochemical Biological Biological Assay Biological Factors Biological Markers Biology Cisplatin Complex DNA Damage DNA Repair DNA biosynthesis DNA repair protein Diagnostic Disease DnaB helicase Drug Targeting Elements Environment Environmental Risk Factor Event Excision Exposure to FDA approved Fluorescence Foundations Future Gene Expression Regulation Genetic Engineering Genus Mycobacterium Goals Granuloma Growth Hypoxia In Situ Infection Intervention Iron Measures Mediating Metabolic Modeling Monitor Mycobacterium tuberculosis Nitric Oxide Organism Oxidation-Reduction Oxidative Stress Oxidoreductase Pathogenesis Performance Pilot Projects Play Post-Translational Protein Processing Post-Translational Regulation Prevention Process Protein Engineering Proteins Pyrococcus abyssi RNA Splicing Rec A Recombinases Recovery Regulation Regulator Genes Reporter Research Role Starvation Sulfur System Testing Therapeutic Transcriptional Activation Tuberculosis Work base catalyst drug development gain of function global health improved in vivo intein macrophage novel oxidative DNA damage promoter public health relevance residence response sensor tool tuberculosis drugs

项目摘要

DESCRIPTION (provided by applicant): The long term goal of this project is to understand the means by which M. tuberculosis (Mtb) regulates its response to the host environment during infection, so that improved intervention strategies and biomarkers can be developed against tuberculosis (TB) disease. Inteins are mobile protein elements that insertionally inactivate the proteins they inhabit. Autocatalytic excision of these inteins results in functional activation of their host proteins, providing an ideal mechanism for rapid post-translational gene regulation. Inteins are found in three Mtb proteins (DnaB, RecA and SufB), all of which have central roles in the process of DNA replication and recovery following exposure to the DNA damaging conditions encountered during mammalian infection. Inteins have been extensively studied at the biochemical level and have been exploited as tools for protein engineering. However, little is known about the biological roles of inteins in Mtb despite recent studies showing that prevention of intein splicing with the FDA approved anti-cancer drug cisplatin inhibits Mtb growth. This pilot project will test the hypothesis that intein splicing is modulated by host-associated environmental conditions and generate a sensitive fluorescence-based gain-of-function reporter system suitable for monitoring Mtb intein splicing within bacteria during infection. Completion of this research plan will provide critical conceptual and technical foundations that will also be needed for future studies addressing the possibility that inteins provide a rapid form of post-translational regulation in Mtb during host infection. This work is of particularly high impact because of the potential for intein splicing to establish new paradigms for gene regulation in Mtb that may control Mtb replication within the host, as well as the demonstrated role of intein splicing as a possible drug target.

描述（由申请人提供）：该项目的长期目标是了解结核分枝杆菌（Mtb）在感染过程中调节其对宿主环境的反应的方式，以便开发改进的针对结核病的干预策略和生物标志物（结核病）疾病。内含肽是可移动的蛋白质元件，可插入地使其所居住的蛋白质失活。这些内含肽的自催化切除导致其宿主蛋白的功能激活，为快速翻译后基因调控提供了理想的机制。内含肽存在于三种 Mtb 蛋白（DnaB、RecA 和 SufB）中，所有这些蛋白在哺乳动物感染期间暴露于 DNA 损伤条件后的 DNA 复制和恢复过程中都发挥着核心作用。内含肽已在生化水平上得到广泛研究，并已被用作蛋白质工程的工具。然而，尽管最近的研究表明，用FDA批准的抗癌药物顺铂来预防内含肽剪接可抑制结核分枝杆菌的生长，但人们对内含肽在结核分枝杆菌中的生物学作用知之甚少。这位飞行员该项目将测试内含肽剪接受宿主相关环境条件调节的假设，并生成敏感的基于荧光的功能获得报告系统，适合监测感染期间细菌内的 Mtb 内含肽剪接。这项研究计划的完成将为未来的研究提供关键的概念和技术基础，以解决内含肽在宿主感染期间在结核分枝杆菌中提供快速翻译后调节形式的可能性。这项工作具有特别高的影响力，因为内含肽剪接有可能在结核分枝杆菌中建立新的基因调控范例，从而控制宿主内的结核分枝杆菌复制，并且内含肽剪接作为可能的药物靶点已被证明发挥作用。