ISOLATION AND ANALYSIS OF HUMAN DNA REPAIR GENES

人类DNA修复基因的分离与分析

• 批准号: 2852944
• 负责人: RANDY J LEGERSKI
• 金额: $ 23.75万
• 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/2852944
• 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.

这一建议的目的是阐明两个新的哺乳动物基因的功能。其中第一个是SNM1，酵母DNA修复基因的哺乳动物同源物。该基因是rad3上位性基因群的成员，已知其特异性参与由链间交联剂产生的损伤的修复。来自酵母研究的证据表明，该基因的表达产物在交联修复的后期发挥作用，影响高分子量DNA的重建。第二个基因与果蝇的北极星基因关系最为密切。在果蝇中对该基因的研究表明，它是一个参与后期染色体分离的有丝分裂因子。北极星的确切功能尚不清楚，据推测它参与了许多过程，包括染色体凝聚、DNA修复或维持姐妹染色单体内聚的因子的溶解，如DNA缠结或蛋白质-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。这些包括作为细胞周期功能的蛋白质表达和定位，蛋白质-蛋白质与新因子或已知因子的相互作用，以及通过反义表达、嵌合寡核苷酸转化和构建纯合子-/-敲除小鼠等几种方法破坏功能。