Probing Metabolic Complexity Using a Bacterial Model System

使用细菌模型系统探测代谢复杂性

• 批准号: 7347533
• 负责人: Jeff M Boyd
• 金额: $ 4.96万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-02-01 至 2009-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7347533
• 关键词: ATP phosphohydrolase; Address; Anabolism; Area; Bacterial Model; Binding; Biochemical; Biochemical Process; Biological Assay; Biological Models; Biology; Cell physiology; Cells; Excision; Face; Free Radicals; Genetic; Genome; Goals; Homeostasis; In Vitro; Internet; Iron; Knowledge; Life; Malignant Neoplasms; Metabolic; Metabolic Pathway; Metabolism; Metals; Open Reading Frames; Organism; Physiological; Physiology; Proteins; Refractory; Research; Role; Salmonella enterica; Series; Starvation; Sulfur; Sum; System; Techniques; Testing; Thiamine; Time; Training; Vitamin, Other; Work; base; career; chemical reaction; genetic analysis; genome sequencing; in vitro Assay; in vivo; repaired; research study; trafficking

A fundamental question in biology is how the integration of metabolic pathways is regulated to produce the robust, but responsive physiology that characterizes sustainable life. Our lack of basic knowledge in these areas is emphasized by the multiple ORFs that remain without functional annotation in diverse genome sequences. Because all living cells face similar challenges in integrating their metabolism, a bacterial system (Salmonella enterica) will be used for simplicity and technical feasibility to complete the outlined objectives. This system will provide training in dissecting a network that can be transferred to more refractory organisms later in my career. Genetic analysis of the system integrated with thiamine biosynthesis has identified a set of five conditional thiamine axotrophs which are defective in Fe-S cluster metabolism. Collectively, these loci appear to be necessary for Fe trafficking, and the repair of oxidatively damaged Fe-S clusters. A series of experiments is outlined to address the specific biochemical function of three of these proteins (ApbE, ApbC, and RseC). The project described herein is divided into two similar and overlapping objectives that will ultimately define the relationship between these and other proteins involved in Fe-S metabolism. Completion of these objectives will require in vivo and in vitro experiments with rigorous application of biochemical, biophysical, genetic, and physiological techniques. The long-term goal of the research presented in this proposal is to contribute to the understanding of metabolic integration, specifically Fe homeostasis. To reach this goal we must increase our knowledge of the biochemical components of metabolism and the connections that exist between them. Such work not only contributes to the rigorous annotation of genomes, but provides the basis for continuing studies on Fe and free radical metabolism. Cellular metabolism is the sum of all of the chemical reactions taking place in a living cell at any given time. Like the internet or the economy, metabolism is a network or system, so small changes, such as the removal of a nuterient, can have profound effects. The objective of the proposed work is to understand the connections between these chemical reactions in order to predict the effects which would be encountered if the system is in some way perturbed (cancer, starvation, desease etc.).

生物学中的一个基本问题是如何调节代谢途径的整合 产生强健但反应灵敏的生理机能，这是可持续生命的特征。我们缺乏基本的 多个 ORF 强调了这些领域的知识，但这些 ORF 仍然没有功能注释 不同的基因组序列。因为所有活细胞在整合新陈代谢方面都面临着类似的挑战， 为了简单性和技术可行性，将使用细菌系统（肠沙门氏菌）来完成 概述的目标。该系统将提供剖析网络的培训，该培训可以转移到更多 在我职业生涯的后期，难治性生物体。 与硫胺素生物合成相结合的系统的遗传分析确定了一组五种 Fe-S簇代谢有缺陷的条件性硫胺素轴突细胞。总的来说，这些位点 似乎对于铁的运输和氧化损伤的铁硫簇的修复是必要的。一系列 概述了实验来解决其中三种蛋白质（ApbE、ApbC、 和 RseC）。本文描述的项目分为两个相似且重叠的目标， 最终定义了这些蛋白质与参与铁硫代谢的其他蛋白质之间的关系。 完成这些目标将需要严格应用体内和体外实验 生物化学、生物物理、遗传和生理学技术。 本提案中提出的研究的长期目标是有助于理解 代谢整合，特别是铁稳态。为了实现这个目标，我们必须增加我们的知识 新陈代谢的生化成分以及它们之间存在的联系。这样的工作不 仅有助于基因组的严格注释，但为继续研究Fe提供了基础 和自由基代谢。 细胞代谢是活细胞中任何时刻发生的所有化学反应的总和 给定时间。就像互联网或经济一样，新陈代谢是一个网络或系统，所以小的变化，例如 营养物质的去除会产生深远的影响。拟议工作的目标是了解 这些化学反应之间的联系，以预测其影响 如果系统受到某种程度的干扰（癌症、饥饿、疾病等），就会遇到这种情况。