Mycobacterium tuberculosis dUTPase as a tool in disease control

结核分枝杆菌 dUTPase 作为疾病控制的工具

• 批准号: 7500520
• 负责人: Judit Toth
• 金额: $ 5.03万
• 依托单位: TERMESZETTUDOMANYI KUTATOKOZPONT
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7500520
• 关键词: Ablation; Active Sites; Address; Anabolism; Bacteria; Bacterial Infections; Basic Science; Biological; C-terminal; Catalysis; Cell Death; Communication; DNA; DNA Maintenance; DNA Repair Enzymes; DNA biosynthesis; Development; Diphosphates; Drug Delivery Systems; Drug resistance; Elements; Engineering; Enzyme Kinetics; Enzymes; Equilibrium; Escherichia coli; Evaluation; Excision; Exhibits; Genes; Genetic; Genus Mycobacterium; Hand; Health; Human; Human Herpesvirus 4; Indium; Investigation; Kinetics; Lead; Measurement; Measures; Metabolic Pathway; Methods; Modeling; Modification; Molecular; Mycobacterium tuberculosis; Organism; Pathway interactions; Pharmaceutical Preparations; Physiological; Plasmodium falciparum; Play; Property; Proteins; Public Health; Range; Rate; Reporter; Research; Role; Solid; Surface; Testing; Thermodynamics; Thymine; Translational Research; Trypanosoma; Tryptophan; Tuberculosis; United States National Institutes of Health; Upper arm; Uracil; Vaccinia; Viral Cancer; Yeasts; abstracting; base; chemical property; concept; dUTP pyrophosphatase; design; disorder control; fighting; gene replacement; homologous recombination; in vivo; inhibitor/antagonist; mutant; mycobacterial; novel; pathogen; prevent; programs; prospective; thymidine 5' -triphosphate; thymidylate; tool

DESCRIPTION (provided by applicant): Project Summary / Abstract dUTPase is the unique enzyme that catalyses the pyrophosphorolysis of dUTP, thus regulating the extent of uracil incorporation into DNA. Massive uracil incorporation may lead to cell death. dUTPase has therefore been recognized as a high-potential drug target in cancer, viral and bacterial disease control. The present proposal focuses on the Mycobacterium tuberculosis (MTB) dUTPase that plays a central role in the mycobacterial dTTP biosynthesis and thus it is likely to be essential for the viability of MTB. MTB is the pathogen that causes tuberculosis, which imposes an increasing global threat with the high-rate emergence of novel multidrug (MDR) and extensively drug-resistant (XDR) strains. Several research agendas (including one at NIH) articulated new measures needed for successful tuberculosis management, which involves intensive research on new drug targets and the development of novel drugs. The present proposal has three Specific Aims all directed towards the evaluation of MTB dUTPase as a valid tool in fighting tuberculosis: 1. Study of the effect of dUTPase functional ablation on the viability of Mycobacterium, 2. Elucidation of the enzymatic mechanism of MTB dUTPase with regards to the mechanistic differences between the human (host) and MTB (pathogen) dUTPases, 3) Determination of the catalytic role of the two structural elements that may be species-specifically targeted in dUTPase. To address the above issues, several transient kinetic and equilibrium enzymological as well as spectroscopical methods will be employed using wild- type and mutant MTB dUTPase enzymes. The physiological effect of functional ablation of dUTPase in Mycobycterium will be investigated in a non-pathogenic Mycobacterium model subjected to dUTPase gene replacement. The expected results of the proposed project will be highly useful in effective species-selective inhibitor design for MTB dUTPase and in the prediction of the in vivo mechanism of such inhibitors. The combination of approaches of this project (cf. Aims 1-3) may serve as a useful concept for the investigation of further potential dUTPase targets such as dUTPases from Plasmodium falciparum; Trypanosoma; vaccinia, herpes and Epstein-Barr viruses. PUBLIC HEALTH RELEVANCE The present proposal focuses on an important DNA repair enzyme called dUTPase of the bacterium causing tuberculosis. Tuberculosis imposes an increasing threat on global health. Therefore several research agendas (including a robust NIH program) articulated new measures needed for successful tuberculosis management, which involves the intensive research of physiological targets for new drugs. The aims of this proposal are directed towards the evaluation of dUTPase as a drug target in tuberculosis control.

描述（由申请人提供）：项目摘要 /摘要DUTPase是催化DUTP的焦磷酸化的独特酶，从而调节尿嘧啶掺入DNA的程度。大量的尿嘧啶掺入可能导致细胞死亡。因此，DUTPase已被公认为是癌症，病毒和细菌疾病控制的高潜力药物靶标。目前的提案着重于结核分枝杆菌（MTB）DUTPase，该dutpase在分枝杆菌DTTP生物合成中起着核心作用，因此对于MTB的可行性可能是必不可少的。 MTB是导致结核病的病原体，随着新型多药（MDR）和广泛的耐药（XDR）菌株的高速出现，施加了日益增加的全球威胁。几项研究议程（包括NIH的一项）阐明了成功的结核病管理所需的新措施，其中涉及对新药物靶标的深入研究和新型药物的发展。本提案的三个具体目的都是针对MTB DUTPase作为对抗结核病的有效工具的三个特定目的：1。研究Dutpase功能消融对分枝杆菌可行性的影响，2。阐明MTB UTPase的酶促机制和MT的机械差异（宿主）（宿主）（宿主）（宿主）（宿主）（宿主）的作用（两个结构元素的催化作用可能是针对Dutpase特异性物种的。为了解决上述问题，将使用野生型和突变的MTB Dutpase酶使用几种短暂的动力学和平衡酶学以及光谱方法。 Dutpase在分枝杆菌中功能消融的生理效果将在接受DUTPase基因替代的非致病分枝杆菌模型中进行研究。该拟议项目的预期结果将在有效的物种选择性抑制剂设计中对MTB Dutpase以及这种抑制剂的体内机制的预测。该项目方法的结合（参见Aims 1-3）可能是研究进一步潜在的dutpase靶标，例如恶性疟原虫的Dutpase靶标的有用概念；锥虫瘤；疫苗，疱疹和爱泼斯坦 - 巴尔病毒。公共卫生相关性本提案着重于一种重要的DNA修复酶，称为dutpase of dutpase，导致结核病。结核病对全球健康构成越来越多的威胁。因此，一些研究议程（包括强大的NIH计划）阐明了成功的结核病管理所需的新措施，其中涉及对新药生理靶标的深入研究。该提案的目的是针对将DUTPase评估为结核病控制的药物靶标。