Interaction of DNA Polymerase alpha with Novel Base-Pairs

DNA 聚合酶 α 与新碱基对的相互作用

基本信息

批准号：
7252551
负责人：
ROBERT D KUCHTA
金额：
$ 3.82万
依托单位：
UNIVERSITY OF COLORADO
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-08-01 至 2008-07-31
项目状态：
已结题

项目摘要

The goal of the proposed studies is to understand how DNA polymerases discriminate between right and wrong dNTPs. In order to better understand how polymerases discriminate between correct and incorrect dNTPs, the interaction of human DNA polymerase alpha with dNTPs will be examined. The specific aims of this proposal are: (1) Develop a complete understanding of how pol alpha discriminates between right and wrong dNTPs. The specific functional groups on a dNTP with which pol alpha interacts will be defined. (2) evelop novel base-pairs that pol alpha replicates with high fidelity and efficiency. To accomplish these goals, a series of dNTP analogs will be synthesized and their interactions with pol alpha determined using steady-state kinetic assays. DNA replicaiton inhibitors comprise a major class of anti-cancer and anti-viral chemotherapeutics, with approximately 2 dozen clinically useful compounds and many more in various stages of development. Thus, a better understanding of the enzymes involved in DNA replication could lead to the development of new chemotherapeutics. A key question regarding DNA and RNA polymerases is how they discriminate between correct and incorrect (d)NTPs. Since DNA polymerase-mediated mutagenesis can lead to cellular transformation, understanding how DNA polymerases discriminate between correct and incorrect dNTPs may lead to a more fundamental understanding of carcinogenesis. This research will be done primarily in the Czech Republic in collaboration with Dr. Michal Hocek as an extension of NIH grant # R01AI059764.

拟议研究的目标是了解 DNA 聚合酶如何区分正确和错误的 dNTP。为了更好地了解聚合酶如何区分正确和不正确的 dNTP，将检查人类 DNA 聚合酶 α 与 dNTP 的相互作用。该提案的具体目标是：（1）全面了解 pol alpha 如何区分正确和错误的 dNTP。将定义 dNTP 上与 pol α 相互作用的特定官能团。 (2) 开发 pol α 以高保真度和效率复制的新碱基对。为了实现这些目标，将合成一系列 dNTP 类似物，并使用稳态动力学测定确定它们与 pol α 的相互作用。 DNA 复制抑制剂是一类主要的抗癌和抗病毒化疗药物，其中有大约 2 打临床有用的化合物以及更多处于不同开发阶段的化合物。因此，更好地了解 DNA 复制所涉及的酶可能会导致新化疗药物的开发。关于 DNA 和 RNA 聚合酶的一个关键问题是它们如何区分正确和错误的 (d)NTP。由于 DNA 聚合酶介导的诱变可导致细胞转化，因此了解 DNA 聚合酶如何区分正确和错误的 dNTP 可能有助于对致癌作用有更基本的了解。这项研究将主要在捷克共和国与 Michal Hocek 博士合作进行，作为 NIH 拨款 # R01AI059764 的延伸。