STUDIES OF DNA POLYMERASES

DNA 聚合酶的研究

• 批准号: 3278172
• 负责人: STUART M LINN
• 金额: $ 21.34万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-06-01 至 1992-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3278172
• 关键词: DNA directed DNA polymerase; DNA repair; Escherichia coli; HeLa cells; affinity chromatography; bacterial DNA; deoxyribonuclease I; dyes; enzyme induction /repression; enzyme reconstitution; enzyme substrate; fibroblasts; gel electrophoresis; human tissue; mitochondrial DNA; monoclonal antibody; nucleic acid sequence; phosphorylation; placenta; protein kinase C; radiotracer; tissue /cell culture

Our long term goals are two-fold: to understand what factors maintain or perturb the fidelity of mammalian DNA polymerases and to understand how these enzymes along with associated DNases take part in DNA excision repair. In the immediate future we propose to study changes which occur in DNA polymerase Alpha as the cell enters a non-cycling state due to population senescence, differentiation or arrest in G-o due to medium exhaustion. We also propose to study how these changes as well as phosphorylation by protein kinase C affect the fidelity and activity levels of DNA polymerase Alpha. Finally, several complex forms of DNA polymerases Alpha, Beta and Delta would be studied in order to learn about factors which affect fidelity. Using very defined damaged DNA and chromatin substrates we propose also to study excision and repair synthesis mediated by several forms of Alpha, Beta and Delta polymerase and by Gamma polymerase, each alone and in conjunction with DNases found to be associated with these enzymes. Finally, we have purified to homogeneity a 220 kilodalton factor which is required for DNA repair synthesis in permeabilized cells and we propose to study the catalytic properties of this factor and its possible relation to DNA polymerase Delta.

我们的长期目标是两个方面：了解哪些因素维持或 扰动哺乳动物DNA聚合酶的保真度，并了解如何了解 这些酶以及相关的DN酶参与DNA切除修复。 在不久的将来，我们建议研究DNA中发生的变化 聚合酶alpha由于人口而进入非周期状态 由于中度精疲力尽而导致G-O的衰老，分化或停滞。 我们 还建议研究这些如何变化以及如何通过 蛋白激酶C会影响DNA聚合酶的保真度和活性水平 阿尔法。 最后，几种复杂形式的DNA聚合酶alpha，beta和 为了了解影响的因素，将研究增量 忠诚。 使用非常定义的受损DNA和染色质底物，我们也建议 研究切除和修复合成由几种形式的α介导的， Beta和Delta聚合酶以及通过伽马聚合酶，单独和 与这些酶有关的DN酶的结合。 最后，我们已经将均匀性纯化为220千达尔顿因素 透化细胞中DNA修复合成所必需的，我们建议 研究该因素的催化特性及其可能的关系 DNA聚合酶三角洲。