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中文摘要
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总结 本研究项目源于我们过去对DNA电荷传输(DNACT)的基础研究 化学.这种化学作用促进了通过DNA双链体的氧化还原化学作用, 敏感地取决于中间碱基对堆叠的完整性。我们现在的目标是应用这个 用于敏感地感测DNA损伤和DNA处理事件的化学, 重要的是,为了确定DNA CT化学如何在细胞内利用的一般特征, 用于远程化学信号。特别是,这些研究可以建立一个新的框架, 考虑到[4Fe 4S]簇在处理DNA的关键酶中的作用。相对 在传感方面,我们将继续开发和应用我们的多路复用平台, 开发用于检测DNA结合蛋白和核酸的新的电化学策略 与癌症相关的标志物。总的来说,我们的多重DNA电化学平台提供了一个 这是一种全新的方法,可以灵敏、良好控制地检测DNA结合事件。我们的整体 目标是开发基于DNA CT化学的高灵敏度传感器, 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;我们将继续我们的研究,以表征至关重要的DNA的氧化还原化学- 处理与复制和DNA修复有关的酶,以确定这种化学反应如何与DNA修复有关。 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芯片表征含有氧化还原活性[4Fe 4S]的DNA加工蛋白 辅因子总的来说,我们迄今为止进行的实验支持DNA CT信号是一种 在DNA加工中具有不同作用的不同蛋白质协调其活性的方法, 团队DNA CT可用于聚合酶传递,并协调对氧化应激的快速反应。 应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转录和复制以及修复,使用AFM测定,遗传学, [4Fe 4S]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能力的生物化学表征,以鉴定新的CT活性蛋白质参与者, 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信号的基本理解和DNA CT的重要考虑 与疾病有关的缺陷。
英文摘要
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.
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DNA Processing Enzymes with [4Fe4S] Clusters for DNA Signaling
BARTON 12-2 PRT
  • 批准号:
    8362342
  • 项目类别:
  • 资助金额:
    $0.14万
  • 财政年份:
    2011
  • 负责人:
    JACQUELINE K BARTON
  • 依托单位:
BARTON 12-2 PRT
  • 批准号:
    8170347
  • 项目类别:
  • 资助金额:
    $0.1万
  • 财政年份:
    2010
  • 负责人:
    JACQUELINE K BARTON
  • 依托单位:
DNA Charge Transport Chemistry & Biology
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