Mycobacterial DNA repair and mutagenesis

分枝杆菌 DNA 修复和诱变

基本信息

批准号：
10619638
负责人：
Michael Stephen Glickman
金额：
$ 66.39万
依托单位：
SLOAN-KETTERING INST CAN RESEARCH
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-02-01 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10619638
关键词：
ATP Hydrolysis ATP phosphohydrolase Actinobacteria class Adenylyl Imidodiphosphate Affect Antibiotic Resistance Antimicrobial Resistance Architecture Award Bacteria Bacterial DNA Binding Biochemical Biological Assay Biological Models C-terminal Cardiolipins Cell membrane Cells Cessation of life Complement Complex Couples Coupling Cryoelectron Microscopy DNA DNA Binding DNA Damage DNA Double Strand Break DNA Interstrand Cross-Link Repair DNA Interstrand Crosslinking DNA Repair DNA Repair Enzymes DNA Repair Pathway DNA glycosylase Data Disease Double Strand Break Repair Drug resistance in tuberculosis Escherichia coli Evolution Excision Family Filament Freezing Funding Genes Genus Mycobacterium Goals Grant Health Human Infection Invaded Knowledge Leprosy Mass Spectrum Analysis Mechanics Membrane Mitomycin C Molecular Conformation Motor Mutagenesis Mutation Mycobacterium Infections Mycobacterium leprae Mycobacterium smegmatis Mycobacterium tuberculosis N-terminal Nucleic Acids Nucleoproteins Pathogenesis Pathway interactions Peptide Hydrolases Phase Phosphatidylinositols Phospholipids Phosphorylation Process Proteins Proteomics Pump Regulation Resected Role SOS Response Serine Single-Stranded DNA Site Structure System Tail Testing Tuberculosis Wit Work arm combat crosslink emerging antibiotic resistance experimental study genetic approach genetic manipulation helicase homologous recombination human pathogen in vivo insight interest lipidomics melting mutant mycobacterial novel novel strategies nuclease pathogen repair strategy repaired response single molecule therapeutic target

项目摘要

Mycobacteria are a genus of the phylum Actinobacteria that includes the human pathogens M. tuberculosis and M. leprae and their avirulent relative M. smegmatis. Human infections with M. tuberculosis (the agent of tuberculosis) and M. leprae (agent of leprosy) cause substantial human suffering. Tuberculosis accounts for ~2 million deaths annually. M. tuberculosis is increasingly antibiotic resistant, solely through the acquisition of mutations in chromosomal genes. The importance of mutagenesis in the evolution of M. tuberculosis antimicrobial resistance, and the related importance of DNA repair pathways in resisting host-inflicted DNA damage, prompts our interest in the mechanisms by which mycobacteria respond to and repair DNA damage. In addition to this important relevance to human health, mycobacteria have emerged as fertile model system to study prokaryotic DNA repair, due to the complexity of the pathways involved and the novel mechanisms that govern these pathways. Our long range goals in this project are to elucidate the DNA repair mechanisms of Mycobacteria. Our work supported by this award has shown that mycobacterial DNA repair differs from the classic E. coli model system with respect to the number of pathway options for the repair of DNA double-strand breaks (DSBs), the roster of DNA repair enzymes, and the regulation of the DNA damage response (DDR). Our premise is that understanding the distinctive features of mycobacterial DNA repair will illuminate the evolution and diversification of repair strategies and suggest new approaches to combat mycobacterial infection and emergence of antibiotic resistance. Our agenda for the next phase of the project focuses on three themes in mycobacterial DNA repair: (i) the mechanism of DSB resection during homologous recombination (HR) by the AdnAB helicase-nuclease; (ii) the role of RecA phosphorylation and RecA interaction with membrane phospholipids in controlling the DNA damage response; and (iii) the structure and repair activities of the DNA helicase Lhr. We propose a combined approach that leverages integrated biochemical, structural, and genetic approaches to understand these DNA repair systems of mycobacteria. Through these studies we will elucidate new mechanisms of DNA repair in mycobacteria, discoveries that will both advance basic knowledge of prokaryotic DNA repair and elucidate pathways relevant to M. tuberculosis drug resistance and pathogenesis.

分枝杆菌是放线菌门的一个属，其中包括人类病原体分枝杆菌。结核分枝杆菌和麻风分枝杆菌及其无毒亲戚耻垢分枝杆菌。人类感染 M. 结核病（结核病病原体）和麻风分枝杆菌（麻风病病原体）导致大量人类死亡痛苦。结核病每年造成约 200 万人死亡。结核分枝杆菌日益增多抗生素耐药性仅通过染色体基因突变获得。这突变在结核分枝杆菌抗菌药物耐药性进化中的重要性，以及 DNA 修复途径在抵抗宿主造成的 DNA 损伤方面的相关重要性，促使我们对分枝杆菌响应和修复 DNA 损伤的机制感兴趣。在除了与人类健康的重要相关性之外，分枝杆菌已成为可繁殖的模型由于所涉及途径的复杂性和控制这些途径的新机制。我们在这个项目中的长期目标是阐明分枝杆菌的 DNA 修复机制。我们的工作得到了该奖项的支持研究表明，分枝杆菌 DNA 修复与经典的大肠杆菌模型系统在以下方面有所不同：修复 DNA 双链断裂 (DSB) 的途径选项数量，名单 DNA 修复酶以及 DNA 损伤反应 (DDR) 的调节。我们的前提是了解分枝杆菌 DNA 修复的独特特征将阐明修复策略的演变和多样化，并提出新的应对方法分枝杆菌感染和抗生素耐药性的出现。我们下一阶段的议程该项目重点关注分枝杆菌 DNA 修复的三个主题：(i) DSB 机制 AdnAB 解旋酶核酸酶在同源重组 (HR) 过程中进行切除； (二) 的作用 RecA 磷酸化以及 RecA 与膜磷脂的相互作用控制 DNA 损害反应； (iii) DNA解旋酶Lhr的结构和修复活性。我们建议利用综合生化、结构和遗传方法的综合方法了解分枝杆菌的 DNA 修复系统。通过这些研究我们将阐明分枝杆菌 DNA 修复的新机制，这些发现都将推进基础研究原核 DNA 修复知识并阐明与结核分枝杆菌药物相关的途径耐药性和发病机制。