Interaction of DNA Polymerase alpha with Base-Pairs

DNA 聚合酶 α 与碱基对的相互作用

• 批准号: 7004867
• 负责人: ROBERT D KUCHTA
• 金额: $ 4.03万
• 依托单位: UNIVERSITY OF COLORADO
• 依托单位国家: 美国
• 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
• 来源: https://reporter.nih.gov/project-details/7004867
• 关键词: Czech Republic; DNA binding protein; DNA directed DNA polymerase; DNA repair; DNA replication; Herpesviridae; chemical synthesis; enzyme activity; molecular shape; polymerization; pyrimidines

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.

拟议的研究的目的是了解DNA聚合酶如何区分对与错误的DNTP。为了更好地了解聚合酶如何区分正确和错误的DNTP，将检查人DNA聚合酶α与DNTP的相互作用。该提案的具体目的是：（1）对Pol Alpha如何区分对与错误的DNTP有一个完整的理解。将定义与Pol Alpha相互作用的DNTP上的特定官能团。 （2）evelop新颖的碱基对，pol alpha以高忠诚和效率复制。为了实现这些目标，将合成一系列DNTP类似物，并与使用稳态动力学测定确定的POLα相互作用。 DNA复制抑制剂包括一类主要的抗癌和抗病毒化学治疗剂，在各个发育阶段中，大约有2二打具有临床上有用的化合物，更多的化合物。因此，对参与DNA复制的酶的更好理解可能导致新的化学疗法的发展。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不正确（d）NTP。由于DNA聚合酶介导的诱变会导致细胞转化，因此了解DNA聚合酶如何区分正确和错误的DNTP可能会导致对癌变的更基本的理解。 这项研究将主要在捷克共和国与Michal Hocek博士合作，作为NIH赠款＃R01AI059764的扩展。