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.

项目概要 甲醛是一种天然存在的代谢物，存在于所有细胞类型中。虽然它与人类有关 疾病，包括痴呆症和糖尿病，它还被证明在有益过程中发挥着关键作用，例如 记忆形成和嘌呤生物合成。在甲基营养细菌中，一碳代谢物如 甲醇可以作为以甲醛为主要中间中间体的途径中的生长底物。 由于其高化学反应性，这些生物体中的甲醛平衡至关重要；然而，他们的 甲醛应激反应系统仍然难以捉摸。 EfgA 和 TtmR 是两个不同的核心角色 调节甲醛抵抗力并破坏甲基营养菌中甲醛稳态的系统 甲基红菌（以前称为甲基杆菌）外扭。 EfgA是新发现的保守甲醛 传感器响应甲醛水平升高而停止生长和翻译。 TtmR 是 MarR 家族 调节许多参与调节、信号传导和应激反应的基因的转录因子，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 对甲醛水平做出反应，以防止不可避免的细胞损伤。具体来说，我们将聘请 基于无偏见测序的方法和实验进化来研究这些机制 系统并定义其监管。甲醛介导的细胞损伤是细胞状态的读数 甲醛稳态；然而，人们对甲醛的体内反应性知之甚少。我们的数据 表明蛋白质损伤是扭伤分枝杆菌细胞毒性的主要原因。我们将使用蛋白质组学 确定甲醛对蛋白质组影响并确定细胞策略的方法 抵消甲醛引起的蛋白质损伤。通过这项工作，我们将利用模型细菌 非常适合维持甲醛稳态，探索甲醛的新兴领域 调节生物学。这项工作的结果将定义重要的细胞过程，并对 有毒代谢物的类似稳态系统。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 有毒代谢物的总体积累，以及代谢物特异性和整体应激反应系统的情况 交叉提供平衡的细胞代谢和生长。