ISOLATION AND ANALYSIS OF HUMAN DNA REPAIR GENES

人类DNA修复基因的分离与分析

• 批准号: 6172384
• 负责人: RANDY J LEGERSKI
• 金额: $ 24.44万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-03-01 至 2002-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6172384
• 关键词: DNA damage; DNA repair; Drosophilidae; antibody neutralization test; antisense nucleic acid; cell cycle; chimeric proteins; crosslink; gene expression; gene targeting; genetically modified animals; human genetic material tag; immunofluorescence technique; immunoprecipitation; laboratory mouse; mutant; oligonucleotides; protein protein interaction; protein structure function; tissue /cell culture; tumor suppressor genes; yeast two hybrid system

The goals of this proposal are to elucidate the function of two novel mammalian genes. The first of these is SNM1, a mammalian homologue of the yeast DNA repair gene. This gene is a member of the rad3 epistasis group and is known to be specifically involved in the repair of lesions produced by interstrand crosslinking agents. Evidence from studies in yeast suggests that the expression product of this gene functions in the later stages of crosslink repair to effect reconstitution of high molecular weight DNA. The second gene is most closely related to the lodestar gene of Drosophila. Studies of this gene in Drosophila suggest that it is a mitotic factor involved in the segregation of chromosomes during anaphase. The exact function of lodestar is not clear, and has been postulated to be involved in a number of processes including chromosomal condensation, DNA repair, or dissolution of the factors that maintain sister chromatid cohesion, such as DNA intertwining or protein-protein interactions. We plan to analyze the function of these genes in mammalian cells by a variety of methods. These include protein expression and localization as a function of the cell cycle, protein-protein interactions with either novel or known factors, and disruption of function through several approaches including antisense expression, conversion by chimeric oligonucleotides, and construction of homozygous -/- knockout mice.

该提案的目标是阐明两个新型哺乳动物基因的功能。 其中的第一个是SNM1，这是酵母DNA修复基因的哺乳动物同源物。 该基因是RAD3上毒基团的成员，已知该基因特别参与了链间交联药物产生的病变的修复。 来自酵母研究的证据表明，该基因在交联修复的后期阶段的表达产物起作用，以实现高分子量DNA的重构。 第二个基因与果蝇的Lodestar基因密切相关。 该基因在果蝇中的研究表明，这是对后期染色体分离的有丝分裂因子。 Lodestar的确切功能尚不清楚，并且已经假定参与了许多过程，包括染色体凝结，DNA修复或溶解维持姊妹染色质剂的因素，例如DNA相互作用或蛋白质 - 蛋白质相互作用。 我们计划通过多种方法分析这些基因在哺乳动物细胞中的功能。 这些包括蛋白质表达和定位是细胞周期的函数，蛋白质 - 蛋白质与新颖或已知因素的相互作用，以及通过多种方法破坏功能，包括反义表达，嵌合寡核苷酸的转化以及纯合子 - / - 敲除小鼠的构建。