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法，以便可以改善干预策略和生物标志物，以针对结核病（TB）疾病发展。 intein是移动蛋白质元素，可在插入它们所居住的蛋白质上失活。这些深度蛋白的自催化切除会导致其宿主蛋白的功能激活，从而为快速翻译后基因调节提供了理想的机制。在三种MTB蛋白（DNAB，RECA和SUFB）中发现了Intein，这些MTB蛋白在暴露于哺乳动物感染期间遇到的DNA损伤条件后所有这些在DNA复制过程中均具有中心作用。在生物化学水平上进行了广泛的研究，并被用作蛋白质工程的工具。然而，尽管最近的研究表明，使用FDA批准的抗癌药物顺铂抑制了MTB的生长，但对Intein在MTB中的生物学作用知之甚少。这个飞行员项目将检验以下假设：内部剪接是由宿主相关的环境条件调节的，并产生基于敏感的荧光赢得的功能记者系统，适用于在感染过程中监测细菌中MTB内素剪接的基础。该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Inteins在宿主感染期间MTB的快速翻译后调节形式的可能性。这项工作的影响尤其很高，因为有可能在MTB中建立可能控制宿主内MTB复制的基因调节的新范例，以及INTEAN剪接作为可能的药物靶标的作用。