Formaldehyde homeostasis and damage repair in a bacterial formaldehyde specialist

细菌甲醛专家的甲醛稳态和损伤修复

• 批准号: 10669782
• 负责人: Jannell V. Bazurto
• 金额: $ 36.93万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-22 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10669782
• 关键词: Anabolism; Bacteria; Biology; Carbon; Cell physiology; Cells; Chemicals; Data; Dementia; Diabetes Mellitus; Equilibrium; Evolution; Family; Formaldehyde; Genes; Growth; Home; Homeostasis; Mediating; Memory; Metabolism; Methanol; Methylobacterium; Methylobacterium extorquens; Modeling; Organism; Pathway interactions; Process; Proteins; Proteome; Proteomics; Purines; Regulation; Resistance; Role; Signal Transduction; Specialist; Stress; System; Toxin; Translations; Work; biological adaptation to stress; cell injury; cell type; cytotoxicity; human disease; in vivo; insight; prevent; repaired; response; sensor; transcription factor

PROJECT SUMMARY Formaldehyde is a naturally occurring metabolite found in all cell types. Although it has been implicated in human disease including dementia and diabetes, it has also shown to have critical roles in beneficial processes such as memory formation and purine biosynthesis. In methylotrophic bacteria, one-carbon metabolites such as methanol can serve as growth substrates in pathways where formaldehyde is an obligate central intermediate. Due to its high chemical reactivity, formaldehyde balance in these organisms is critical; however, their formaldehyde stress response systems have remained elusive. EfgA and TtmR are central players of two distinct systems that modulate formaldehyde resistance and disrupt formaldehyde homeostasis in the methylotroph Methylorubrum (formerly Methylobacterium) extorquens. EfgA is a newly identified conserved formaldehyde sensor that halts growth and translation in response to elevated formaldehyde levels. TtmR is a MarR-family transcription factor that regulates many genes involved in regulation, signaling, and stress response, including efgA. Our work will characterize the EfgA and TtmR homeostasis systems to understand how cells sense and respond to formaldehyde levels to prevent otherwise inevitable cellular damage. Specifically, we will employ unbiased sequencing-based approaches and experimental evolution to home in on the mechanisms of these systems and define their regulation. Formaldehyde-mediated cellular damage is a readout of the status of formaldehyde homeostasis; however, the in vivo reactivity of formaldehyde is poorly understood. Our data suggests that protein damage is the predominant cause of cytotoxicity in M. extorquens. We will use proteomics approaches to define the impact of formaldehyde on the proteome and identify cellular strategies for counteracting formaldehyde-induced protein damage. Through this work, we will leverage a model bacterium that is well adapted to maintain formaldehyde homeostasis to explore the burgeoning field of formaldehyde regulatory biology. The results from this work will define essential cellular processes and has implications for analogous homeostasis systems for toxic metabolites. We envision this work will have substantial impacts on the understanding of how cells sense and regulate formaldehyde levels, how cells navigate and avoid accumulation of toxic metabolites generally, and how metabolite-specific and global systems of stress response intersect to provide balanced cellular metabolism and growth.

项目摘要 甲醛是一种天然存在的代谢物，存在于所有细胞类型中。尽管它与人类的 它还被证明在有益的过程中发挥关键作用，例如 记忆形成和嘌呤生物合成。在甲基营养型细菌中，一碳代谢物如 甲醇可以在其中甲醛是专性中心中间体的途径中用作生长底物。 由于其高化学反应性，甲醛在这些生物体中的平衡是至关重要的;然而， 甲醛应激反应系统仍然难以捉摸。EfgA和TtmR是两种不同的 甲基营养菌中调节甲醛抗性和破坏甲醛稳态的系统 扭脱甲基红杆菌（Methylorubrum）。EfgA是一种新发现的保守甲醛 传感器，停止生长和翻译，以响应甲醛水平升高。TtmR是一个MarR家族 一种转录因子，调节许多参与调控、信号传导和应激反应的基因，包括 efgA.我们的工作将表征EfgA和TtmR稳态系统，以了解细胞如何感知和 对甲醛含量作出反应，以防止不可避免的细胞损伤。具体来说，我们将采用 无偏见的测序为基础的方法和实验进化，以家庭在这些机制 系统并定义其规则。甲醛介导的细胞损伤是一个状态的读数， 甲醛稳态;然而，甲醛的体内反应性知之甚少。我们的数据 提示蛋白质损伤是M细胞毒性的主要原因。勒索。我们将使用蛋白质组学 确定甲醛对蛋白质组的影响并确定细胞策略的方法 抵消甲酰胺诱导的蛋白质损伤。通过这项工作，我们将利用一种模型细菌， 这是很好地适应维持甲醛稳态探索甲醛的新兴领域 调节生物学。这项工作的结果将定义基本的细胞过程，并对 有毒代谢物的类似体内平衡系统。我们预计这项工作将对 了解细胞如何感知和调节甲醛水平，细胞如何导航和避免 一般有毒代谢物的积累，以及代谢物特异性和全球系统的应激反应 交叉以提供平衡细胞代谢和生长。