DNA Replication Fidelity

工作概述：该项目的长期目标是了解多蛋白DNA复制和修复复合物合成DNA的保真度。今年，我们在三个方面取得了进展。我们测定了酵母DNA聚合酶epsilon合成DNA的保真度，epsilon是负责复制真核生物基因组的两种主要聚合酶之一。为了研究DNA聚合酶epsilon在细胞中的作用，我们确定了DNA聚合酶epsilon活性位点的点突变对其他复制蛋白存在多种缺陷的酵母菌株突变率的影响。我们还研究了酵母DNA聚合酶delta和epsilon在翻译合成胸腺嘧啶二聚体时的聚合和纠错中的作用，胸腺嘧啶二聚体是暴露在阳光下产生的光产物，负责致突变的潜在皮肤癌易感性。

Summary of Work: The long-term goal of this project is to understand the fidelity of DNA synthesis by multiprotein DNA replication and repair complexes. This year, progress was made in three areas. We determined the fidelity of DNA synthesis by yeast DNA polymerase epsilon, one of the two major polymerases responsible for replicating the eukaryotic genome. To investigate the role of DNA polymerase epsilon in cells, we determined the influence of a point mutation in the active site of DNA polymerase epsilon on mutation rates in yeast strains with a variety of defects in other replication proteins. We also investigated the roles of yeast DNA polymerases delta and epsilon in polymerization and error correction during translesion synthesis past a thymine dimer, a photoproduct resulting from exposure to sunlight and responsible for the mutagenesis underlying susceptibility to skin cancer. Background: DNA polymerases do not work alone in cells, but function within interconnected DNA processing pathways that require many other proteins to replicate DNA and to repair various types of environmental DNA damage. Some of these proteins determine which substrates a DNA polymerase can copy and/or influence the efficiency and fidelity with which DNA synthesis occurs. Building on our structure-function studies of purified DNA polymerases, the goal of this project is to understand the fidelity of DNA synthesis by multi-protein DNA replication and DNA repair complexes, and to understand how perturbing these complexes may result in genome instability associated with environmental disease. A critical component of this effort is to connect in vi