Probing Metabolic Complexity Using a Bacterial Model System

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 这些领域的知识是由多个ORF强调的， 不同的基因组序列。因为所有的活细胞在整合它们的新陈代谢时都面临着类似的挑战， 为了简单和技术可行性，将使用细菌系统（肠道沙门氏菌）来完成 概述的目标。该系统将提供解剖网络的培训，该网络可以转移到更多 在我的职业生涯后期， 与硫胺素生物合成整合的系统的遗传分析已经确定了一组五个 条件性硫胺素轴养体，其在Fe-S簇代谢中有缺陷。总的来说，这些位点 似乎是必要的铁运输，和氧化损伤的Fe-S簇的修复。一系列 概述了实验以阐明这些蛋白质中的三种（ApbE，ApbC， 和RseC）。本文所述的项目分为两个类似和重叠的目标， 最终确定这些蛋白质和其他参与Fe-S代谢的蛋白质之间的关系。 这些目标的完成将需要在体内和体外实验中严格应用 生物化学、生物物理学、遗传学和生理学技术。 本提案中提出的研究的长期目标是促进对 代谢整合，特别是铁稳态。为了实现这一目标，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的联系。这样的工作不 这不仅有助于基因组的严格注释，而且为继续研究Fe提供了基础 和自由基代谢。 细胞代谢是在任何时间在活细胞中发生的所有化学反应的总和。 给你时间就像互联网或经济一样，新陈代谢是一个网络或系统，因此， 去除营养素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，以预测这些化学反应的影响， 如果系统以某种方式受到干扰（癌症，饥饿，疾病等），就会遇到这种情况。

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.).