DNA Sensing and Signaling

DNA 传感和信号转导

• 批准号: 9891857
• 负责人: JACQUELINE K BARTON
• 金额: $ 60.3万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9891857
• 关键词: Anaerobic Bacteria; Base Pairing; Biochemical; Biological Assay; Cells; Characteristics; Charge; Chemicals; Chemistry; DNA; DNA Binding; DNA Microarray Chip; DNA Repair; DNA biosynthesis; DNA lesion; DNA-Binding Proteins; Detection; Development; Disease; Electrochemistry; Enzymes; Event; Foundations; Genetic; Genetic Transcription; Genome; Goals; Hand; Malignant Neoplasms; Monitor; Mutation; Nature; Nucleic Acids; Oxidation-Reduction; Oxidative Stress; Polymerase; Process; Proteins; Research; Role; STEM program; Signal Transduction; Spectrum Analysis; Work; base; cancer biomarkers; cofactor; experimental study; mutant; novel strategies; repaired; response; sensor

Summary This research program stems from our past fundamental studies of DNA charge transport (DNA CT) chemistry. This chemistry facilitates redox chemistry at a distance through the DNA duplex and sensitively depends upon the integrity of the intervening base pair stack. Our goal now is to apply this chemistry for the sensitive sensing of DNA lesions and DNA processing events electrically and, importantly, to determine general characteristics of how DNA CT chemistry is utilized within the cell for long range chemical signaling. In particular, these studies may establish a new framework for considering the role of [4Fe4S] clusters in critically important enzymes that process DNA. With respect to sensing, we will continue in the development and application of our multiplexed platform in developing new electrochemical strategies for the detection of DNA-binding proteins and nucleic acid markers associated with cancer. In general, our multiplexed DNA electrochemical platform offers a completely new approach to sensitive, well-controlled detection of DNA-binding events. Our overall goal is the development of highly sensitive sensors based on DNA CT chemistry that can be used to monitor biomolecules in a single cell. Significantly, our goal also is to understand how Nature may use DNA CT; we will continue our studies to characterize the redox chemistry of critically important DNA- processing enzymes involved in replication and DNA repair to determine how this chemistry may be utilized for long range signaling across the genome. We will use DNA electrochemistry anaerobically using multiplexed DNA chips to characterize DNA-processing proteins containing redox-active [4Fe4S] cofactors. Overall, experiments we have carried out thus far provide support that DNA CT signaling is a means for diverse proteins with different roles in DNA processing to coordinate their activities as a team. DNA CT may be used for polymerase hand-offs and to coordinate a rapid response to oxidative stress. We will build upon this foundation by exploring DNA CT signaling among protein teams involved in DNA transcription and replication as well as repair, using AFM assays, genetics, and spectroscopy on [4Fe4S] proteins and mutants. We aim to use our expertise in the electrochemical and biochemical characterization of DNA CT proficiency to identify new CT-active protein players and illuminate the DNA CT signaling networks in which they participate. This work will contribute both to our fundamental understanding of protein/DNA signaling and the important consideration of DNA CT deficiencies associated with disease.

概括 该研究项目源于我们过去对 DNA 电荷传输 (DNA CT) 的基础研究 化学。这种化学反应通过 DNA 双链体促进远距离的氧化还原化学反应 敏感地取决于中间碱基对堆栈的完整性。我们现在的目标是应用这个 用于对 DNA 损伤和 DNA 加工事件进行电敏感传感的化学方法， 重要的是，确定 DNA CT 化学在细胞内如何利用的一般特征 用于长距离化学信号传导。特别是，这些研究可能会建立一个新的框架 考虑 [4Fe4S] 簇在处理 DNA 的极其重要的酶中的作用。带着尊重 到传感，我们将继续开发和应用我们的多路复用平台 开发新的电化学策略来检测 DNA 结合蛋白和核酸 与癌症相关的标志物。一般来说，我们的多重 DNA 电化学平台提供了 一种灵敏、控制良好的 DNA 结合事件检测的全新方法。我们的整体 目标是开发基于 DNA CT 化学的高灵敏度传感器，可用于 监测单个细胞中的生物分子。值得注意的是，我们的目标也是了解大自然如何利用 DNA CT；我们将继续研究来表征极其重要的 DNA 的氧化还原化学- 处理参与复制和 DNA 修复的酶以确定这种化学反应如何进行 用于跨基因组的长距离信号传导。我们将在厌氧条件下使用 DNA 电化学 使用多重 DNA 芯片来表征含有氧化还原活性 [4Fe4S] 的 DNA 加工蛋白 辅助因子。总体而言，我们迄今为止进行的实验支持 DNA CT 信号传导是一种 意味着在 DNA 加工中具有不同作用的不同蛋白质能够协调其活动 团队。 DNA CT 可用于聚合酶交接并协调对氧化的快速反应 压力。我们将在此基础上探索蛋白质团队之间的 DNA CT 信号传导 参与 DNA 转录、复制以及修复，使用 AFM 分析、遗传学和 [4Fe4S] 蛋白质和突变体的光谱。我们的目标是利用我们在电化学和 DNA CT 熟练程度的生化表征，以识别新的 CT 活性蛋白参与者和 阐明它们参与的 DNA CT 信号网络。这项工作将有助于 我们对蛋白质/DNA 信号传导的基本了解以及 DNA CT 的重要考虑 与疾病相关的缺陷。