Probing Metabolic Complexity Using a Bacterial Model System

生物学中的一个基本问题是新陈代谢途径的整合是如何调节的 产生强大但反应灵敏的生理学，这是可持续生活的特征。我们对基础设施的缺乏 中没有功能注释的多个ORF强调了这些领域的知识 不同的基因组序列。因为所有活着的细胞在整合新陈代谢方面都面临着相似的挑战， 为了简便和技术上的可行性，将使用细菌系统(沙门氏菌)来完成 勾勒出目标。该系统将提供解剖网络的培训，该网络可以转移到更多 在我职业生涯的后期。 结合硫胺素生物合成的系统的遗传分析已经确定了一组五个 铁-S簇代谢缺陷的条件性硫胺素促轴营养细胞。总的来说，这些基因座 似乎对铁的运输是必要的，并修复氧化损伤的铁-S团簇。一系列 实验概述了这些蛋白质中的三种(ApbE，APBC， 和RseC)。这里描述的项目被分成两个相似且重叠的目标，它们将 最终确定这些蛋白质与其他参与铁-S代谢的蛋白质之间的关系。 完成这些目标将需要体内和体外实验，并严格应用 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，我们必须增加我们对 新陈代谢的生化成分以及它们之间存在的联系。这样的工作不是 仅有助于对基因组的严格注释，但为继续研究铁提供了基础 和自由基代谢。 细胞新陈代谢是活细胞中发生的所有化学反应的总和 给你点时间。就像互联网或经济一样，新陈代谢是一个网络或系统，所以一些小的变化，比如 一种营养物的去除，可能会产生深远的影响。拟议工作的目标是了解 这些化学反应之间的联系，以便预测 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.).