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Base Pair Opening in the Origin of DNA Replication

DNA 复制起源中的碱基对打开

基本信息

批准号：
6457347
负责人：
IRINA M RUSSU
金额：
$ 15.1万
依托单位：
WESLEYAN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-04-01 至 2006-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6457347
关键词：
DNA replication origin Escherichia coli bioenergetics chemical stability chromosomes cytogenetics genetic mapping high performance liquid chromatography hydrogen ions nuclear magnetic resonance spectroscopy nucleic acid sequence nucleobase replicon

项目摘要

DESCRIPTION: (provided by applicant) Transmission of genetic information in living cells occurs by replication of the DNA into two identical copies. This process is initiated at specific sites on the chromosomes, where the double helix opens and the parental strands separate in order to allow the entry and the action of the replication machinery. These specific sites where replication begins have acquired through evolution unique energetic features that allow them to function as DNA Unwinding Elements (DUE). The research proposed in this application aims at mapping the energetics of opening of each base pair in the DNA Unwinding Elements of a bacterial chromosome. The method of choice is nuclear magnetic resonance (NMR) spectroscopy. 1/H and 15/N NMR will be used to observe the imino protons in each Watson-Crick base pair of three DNA duplexes containing the DUE sites of interest. The rates of exchange of these protons with solvent will be measured by NMR as a function of temperature and concentration of exchange catalyst. The exchange rates will define the kinetics and energetics of opening of each base pair in these DUE sites. These high-resolution energetic maps will contribute to the understanding of the fundamental relationships between base sequence and local stability in DNA double helices.

描述：（由申请人提供）遗传信息的传输活细胞是通过将 DNA 复制成两个相同的拷贝而产生的。这该过程在染色体上的特定位点启动，其中双螺旋打开，亲本链分离，以允许进入和复制机器的动作。这些特定的站点进行复制开始通过进化获得了独特的能量特征，使它们充当 DNA 解旋元件 (DUE)。本文提出的研究应用程序旨在绘制每个碱基对打开的能量图细菌染色体的 DNA 解旋元件。选择的方法是核磁共振（NMR）光谱。 1/H 和 15/N NMR 将用于观察三个 DNA 双链体的每个 Watson-Crick 碱基对中的亚氨基质子包含感兴趣的 DUE 位点。这些质子的交换率与溶剂的关系将通过 NMR 测量作为温度和交换催化剂的浓度。汇率将决定动力学以及这些 DUE 位点中每个碱基对打开的能量。这些高分辨率能量图将有助于理解 DNA 碱基序列与局部稳定性之间的基本关系双螺旋。