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: 7645141
• 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/7645141
• 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; Elements; Engineering; Enzyme Kinetics; Enzymes; Equilibrium; Escherichia coli; Evaluation; Excision; Exhibits; Extreme drug resistant tuberculosis; 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; 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; dUTP pyrophosphatase; design; disorder control; drug candidate; fighting; gene replacement; homologous recombination; in vivo; inhibitor/antagonist; mutant; mycobacterial; novel; pathogen; prevent; programs; prospective; public health relevance; 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 是一种独特的酶，可催化 dUTP 的焦磷酸解，从而调节尿嘧啶掺入 DNA 的程度。大量尿嘧啶掺入可能导致细胞死亡。因此，dUTPase 被认为是癌症、病毒和细菌疾病控制中的高潜力药物靶点。本提案重点关注结核分枝杆菌 (MTB) dUTP 酶，该酶在分枝杆菌 dTTP 生物合成中发挥核心作用，因此可能对 MTB 的生存至关重要。 MTB 是导致结核病的病原体，随着新型多药 (MDR) 和广泛耐药 (XDR) 菌株的大量出现，结核病对全球的威胁日益严重。几个研究议程（包括美国国立卫生研究院的一个）阐明了成功结核病管理所需的新措施，其中涉及对新药物靶点的深入研究和新药的开发。本提案有三个具体目标，均旨在评估 MTB dUTPase 作为抗击结核病的有效工具：1. 研究 dUTPase 功能消融对分枝杆菌活力的影响，2. 阐明 MTB dUTPase 的酶促机制，了解人类（宿主）和 MTB（病原体）dUTPase 之间的机制差异，3）确定 dUTPase 中可能特异性靶向的两个结构元件的催化作用。为了解决上述问题，将使用野生型和突变型 MTB dUTPase 酶来采用几种瞬时动力学和平衡酶学以及光谱方法。将在经过 dUTPase 基因替换的非致病性分枝杆菌模型中研究分枝杆菌中 dUTPase 功能性消除的生理效应。该项目的预期结果对于 MTB dUTPase 的有效物种选择性抑制剂设计以及此类抑制剂的体内机制预测非常有用。该项目的方法组合（参见目标 1-3）可以作为研究进一步潜在 dUTPase 靶标（例如来自恶性疟原虫的 dUTPase）的有用概念；锥虫；痘苗病毒、疱疹病毒和 Epstein-Barr 病毒。公共卫生相关性 本提案重点关注引起结核病的细菌中一种称为 dUTPase 的重要 DNA 修复酶。结核病对全球健康造成越来越大的威胁。因此，几个研究议程（包括强大的 NIH 计划）阐明了成功结核病管理所需的新措施，其中涉及对新药生理目标的深入研究。该提案的目的是评估 dUTPase 作为结核病控制药物靶点的效果。