Legerski R J, Penkala J E, Peterson C A, Wright D A
Department of Genetics, University of Texas M. D. Anderson Hospital and Tumor Institute, Houston 77030.
Mol Cell Biol. 1987 Dec;7(12):4317-23. doi: 10.1128/mcb.7.12.4317-4323.1987.
We characterized a DNA repair system in frog oocytes by comicroinjection of UV-irradiated pBR322 DNA and radiolabeled nucleotides. Repair synthesis was monitored by incorporation of label into recovered pBR322 DNA and by a novel method in which the removal of UV photoproducts was determined from the shift of DNA topoisomers that occurs during gel electrophoresis upon repair of these lesions. We investigated the effects of several drugs in the oocyte system and found that although novobiocin, an inhibitor of topoisomerase II, was an effective inhibitor of repair, VM-26, another inhibitor of topoisomerase II, was not. In addition, the topoisomerase I inhibitor camptothecin had no effect on repair in this system. Finally, circular DNA (either supercoiled or nicked circular) was repaired at least 50 times more rapidly than linear DNA.
我们通过共显微注射紫外线照射的pBR322 DNA和放射性标记的核苷酸，对蛙卵母细胞中的DNA修复系统进行了表征。通过将标记物掺入回收的pBR322 DNA中以及一种新方法来监测修复合成，该新方法是根据在这些损伤修复过程中凝胶电泳时发生的DNA拓扑异构体的变化来确定紫外线光产物的去除情况。我们研究了几种药物在卵母细胞系统中的作用，发现虽然拓扑异构酶II抑制剂新生霉素是修复的有效抑制剂，但另一种拓扑异构酶II抑制剂VM - 26却不是。此外，拓扑异构酶I抑制剂喜树碱在该系统中对修复没有影响。最后，环状DNA（超螺旋或带切口的环状）的修复速度比线性DNA至少快50倍。
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