Regulation of C1 Metabolism in Methylotrophs

甲基营养菌 C1 代谢的调节

• 批准号: 7025039
• 负责人: Mary E Lidstrom
• 金额: $ 30.16万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-08-01 至 2008-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7025039
• 关键词: 1 carbon compound; Methanobacteriaceae; bacterial genetics; biochemical evolution; enzyme activity; formaldehyde; formates; gene expression; gene targeting; genetic regulation; methane; microarray technology; microorganism culture; microorganism metabolism; molecular cloning; oxidation; proteomics; transfection

DESCRIPTION (provided by applicant): The production, interconversion, and transfer of C1 units is an important metabolic system in all of biology. Methylotrophs are microorganisms capable of growth on C1 compounds as sole carbon and energy sources, and methylotrophy can be viewed as a specialized version of the C1 metabolism found in all organisms. A distinguishing feature of methylotrophic metabolism is the generation and consumption of formaldehyde as a central intermediate, the starting point for all of metabolism. We have gained major new insights into the pathways that consume formaldehyde in Methylobacterium extorquens AM1, and have developed a model for how the cell controls formaldehyde flux to achieve a dynamic balance of carbon and energy metabolism, avoiding formaldehyde toxicity. In this project, we will focus on the three pathways, or modules, that our working model predict are central to understanding flux of formaldehyde and energy metabolism, the H4MPT pathway, the H4F pathway, and the 3 formate dehydrogenases. We propose to begin to test our conceptual model of formaldehyde-related central metabolism in methylotrophy using a combination of biochemical, genetic, genomic, and computational approaches, focused initially on understanding how the cell responds to changes in the formaldehyde production rate. This complex system has two fundamental circuits that will be analyzed, the genetic circuit, consisting of the transcriptional and translational elements and the associated signaling components, with the output being transcripts and proteins, and the metabolic circuit, consisting of enzymes, cofactors, intermediates, and the associated signaling compounds, with the output being metabolic flux. Because of the difficulty of measuring all of the components and their characteristics, we will take a modular approach and measure outputs for each of the modules, integrating the results to create a systems-level understanding of response and resultant effects. The specific aims are: 1. Analyze the output of the genetic circuit with microarrays and proteomics. 2. Analyze the output of the metabolic circuit with enzyme assays, metabolite measurements, and direct flux measurements. 3. Integrate the results using computational models that correlate the functioning of the genetic circuit and the metabolic circuit. The result of this study will be a systems-level understanding of formaldehyde metabolism in methylotrophy. These approaches will provide a model for functional genomics at the physiological level, and will create a platform for future studies of the interaction between normal and stressed metabolism, and the mechanistic understanding of the interplay between genetic and metabolic circuits.

描述（由申请人提供）：C1单位的产生，相互转换和转移是所有生物学中重要的代谢系统。甲基化营养体是一种能够以C1化合物为唯一碳源和能量源生长的微生物，甲基化营养体可以被视为所有生物中发现的C1代谢的特殊版本。甲基营养代谢的一个显著特征是甲醛作为中心中间体的产生和消耗，甲醛是所有代谢的起点。我们对敲诈甲基杆菌AM1消耗甲醛的途径有了重要的新见解，并建立了细胞如何控制甲醛通量以实现碳和能量代谢的动态平衡，避免甲醛毒性的模型。在这个项目中，我们将重点关注我们的工作模型预测的三个途径或模块，即H4MPT途径、H4F途径和3种甲酸脱氢酶，它们是理解甲醛通量和能量代谢的核心。我们建议开始使用生化、遗传、基因组和计算方法的组合来测试甲基化过程中甲醛相关中枢代谢的概念模型，最初的重点是了解细胞如何对甲醛产生速率的变化做出反应。这个复杂的系统有两个将被分析的基本回路，一个是遗传回路，由转录和翻译元件以及相关的信号元件组成，输出是转录物和蛋白质，另一个是代谢回路，由酶、辅因子、中间体和相关的信号化合物组成，输出是代谢通量。由于测量所有组件及其特性的困难，我们将采用模块化方法并测量每个模块的输出，整合结果以创建对响应和结果效果的系统级理解。具体目标是：1。用微阵列和蛋白质组学分析遗传电路的输出。2. 用酶分析、代谢物测量和直接通量测量来分析代谢回路的输出。3. 使用计算模型将结果整合起来，这些模型将遗传回路和代谢回路的功能联系起来。这项研究的结果将是对甲基化过程中甲醛代谢的系统级理解。这些方法将为生理水平上的功能基因组学提供一个模型，并将为未来研究正常和应激代谢之间的相互作用以及遗传和代谢回路之间相互作用的机制理解创造一个平台。