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 该研究计划源于我们过去对DNA电荷运输（DNA CT）的基本研究 化学。这种化学促进了通过DNA双链体的距离氧化还原化学反应 敏感地取决于中间基对堆栈的完整性。我们现在的目标是应用 对DNA病变和DNA处理事件的敏感感应的化学性质，以及 重要的是，要确定细胞中如何利用DNA CT化学的一般特征 用于远程化学信号。特别是，这些研究可能为 考虑[4FE4S]簇在处理DNA的至关重要的酶中的作用。尊重 感知，我们将继续在我们的多路复用平台的开发和应用中 开发新的电化学策略来检测DNA结合蛋白和核酸 与癌症相关的标记。通常，我们的多路复用DNA电化学平台提供了 全新的方法来敏感，控制良好的DNA结合事件。我们的整体 目标是基于DNA CT化学的高度敏感传感器的开发，该传感器可用于 监测单个细胞中的生物分子。重要的是，我们的目标也是了解大自然如何使用 DNA CT;我们将继续研究以表征至关重要的DNA-的氧化还原化学 处理复制和DNA修复的加工酶，以确定这种化学的方式 用于跨基因组的远程信号传导。我们将在厌氧上使用DNA电化学 使用多路复用的DNA芯片来表征含有氧化还原活性的DNA处理蛋白[4FE4S] 辅因子。总体而言，迄今为止我们已经进行的实验提供了支持DNA CT信号是一个 用于在DNA加工中具有不同作用的多种蛋白质的手段，以协调其活性作为一个 团队。 DNA CT可用于聚合酶交接并协调对氧化的快速响应 压力。我们将通过探索蛋白质团队中的DNA CT信号来建立基础 参与DNA转录和复制以及维修，使用AFM分析，遗传学和 [4FE4S]蛋白质和突变体的光谱法。我们的目标是在电化学和 DNA CT能力的生化表征，以鉴定新的CT活性蛋白参与者和 照亮他们参与的DNA CT信号网络。这项工作将兼而有之 我们对蛋白质/DNA信号传导的基本理解和DNA CT的重要考虑 与疾病相关的缺陷。