DNA Sensing and Signaling

摘要 这项研究计划源于我们过去对DNA电荷传输(DNA CT)的基础研究 化学反应。这种化学作用促进了远距离的氧化还原化学通过DNA双链和 敏感地依赖于中间碱基对堆叠的完整性。我们现在的目标是应用这一点 用于敏感感应DNA损伤和DNA处理事件的化学， 重要的是，为了确定DNA CT化学如何在细胞内被利用的一般特征 用于远距离化学信号。特别是，这些研究可能会建立一个新的框架 考虑到[4Fe4S]团簇在处理DNA的关键酶中的作用。怀着敬意 为了感知，我们将继续开发和应用我们的多路复用平台在 发展检测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，我们的多路DNA电化学平台提供了 一种全新的灵敏、可控的DNA结合事件检测方法。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自然如何利用 DNA CT；我们将继续研究至关重要的DNA的氧化还原化学- 处理参与复制和DNA修复的酶，以确定这种化学作用可能是如何 用于整个基因组的远程信号传递。我们将把DNA电化学用于厌氧 利用多重DNA芯片表征含有氧化还原活性的DNA加工蛋白[4Fe4S] 辅因。总体而言，我们到目前为止进行的实验支持了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作为 一队。DNA CT可用于聚合酶交接和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 参与DNA转录和复制以及修复，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的生化特征用于识别新的CT活性蛋白和 说明他们参与的DNA CT信号网络。这项工作将有助于 我们对蛋白质/DNA信号的基本认识和DNA CT的重要考虑 与疾病相关的缺陷。

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.