Wood R D, Coverley D
Imperial Cancer Research Fund, Clare Hall Laboratories, South Mimms, Herts, UK.
Bioessays. 1991 Sep;13(9):447-53. doi: 10.1002/bies.950130904.
The many genetic complementation groups of DNA excision-repair defective mammalian cells indicate the considerable complexity of the excision repair process. The cloning of several repair genes is taking the field a step closer to mechanistic studies of the actions and interactions of repair proteins. Early biochemical studies of mammalian DNA repair in vitro are now at hand. Repair synthesis in damaged DNA can be monitored by following the incorporation of radiolabelled nucleotides. Synthesis is carried out by mammalian cell extracts and is defective in extracts from cell lines derived from individuals with the excision-repair disorder xeroderma pigmentosum. Biochemical complementation of the defective extracts can be used to purify repair proteins. Repair of damage caused by agents including ultraviolet irradiation, psoralens, and platinating compounds has been observed. Neutralising antibodies against the human single-stranded DNA binding protein (HSSB) have demonstrated a requirement for this protein in DNA excision repair as well as in DNA replication.
DNA切除修复缺陷的哺乳动物细胞的众多基因互补组表明切除修复过程相当复杂。几个修复基因的克隆使该领域在对修复蛋白的作用和相互作用进行机制研究方面又迈进了一步。哺乳动物DNA修复的早期体外生化研究现已开展。受损DNA中的修复合成可通过追踪放射性标记核苷酸的掺入来监测。合成由哺乳动物细胞提取物进行,并且在源自患有切除修复障碍——着色性干皮病个体的细胞系提取物中存在缺陷。缺陷提取物的生化互补可用于纯化修复蛋白。已观察到对包括紫外线照射、补骨脂素和铂化合物在内的试剂所造成损伤的修复。针对人类单链DNA结合蛋白(HSSB)的中和抗体已证明该蛋白在DNA切除修复以及DNA复制中是必需的。