Lommel L, Hanawalt P C
Department of Biological Sciences, Stanford University, CA 94305-5020.
Mutat Res. 1991 Sep;255(2):183-91. doi: 10.1016/0921-8777(91)90052-q.
We examined removal of cyclobutane pyrimidine dimers (CPDs) from the dihydrofolate reductase (DHFR) gene in ultraviolet-irradiated Chinese hamster ovary (CHO) UV61 and UV5 cells. The sensitivity of UV61 cells to UV-irradiation is intermediate between that of the parental CHO cells and that of mutants such as UV5 that are highly defective in excision repair. UV61 cells have been characterized as having normal repair of pyrimidine(6-4)pyrimidone photoproducts (6-4 PPs) but no detectable removal of CPDs from the genome overall. We find that the extent of removal of CPDs from the DHFR gene in UV61 cells is intermediate between that of the parental CHO cells and that of the UV5 mutant, and the observed repair appears to be confined to the transcribed strand. We detected no removal of CPDs from the DHFR gene in UV5 cells. Our findings in UV61 cells demonstrate a correlation between survival after UV-irradiation and CPD repair in an expressed gene in a cell line with moderate UV-sensitivity and yet no apparent removal of CPDs from the genome as a whole. We have thus demonstrated that overall repair measurements can be misleading. Our results have implications for the determination of the relative biological importance of the CPD and the 6-4 PP, and they further support the hypothesis that removal of CPDs from transcriptionally active DNA is crucial for UV-resistance.
我们检测了紫外线照射的中国仓鼠卵巢(CHO)UV61和UV5细胞中二氢叶酸还原酶(DHFR)基因中环丁烷嘧啶二聚体(CPD)的去除情况。UV61细胞对紫外线照射的敏感性介于亲代CHO细胞和诸如UV5等切除修复高度缺陷的突变体之间。UV61细胞的特征是嘧啶(6-4)嘧啶酮光产物(6-4PPs)修复正常,但总体上基因组中未检测到CPD的去除。我们发现,UV61细胞中DHFR基因CPD的去除程度介于亲代CHO细胞和UV5突变体之间,并且观察到的修复似乎局限于转录链。我们在UV5细胞中未检测到DHFR基因中CPD的去除。我们在UV61细胞中的发现表明,在具有中等紫外线敏感性且基因组整体上没有明显CPD去除的细胞系中,紫外线照射后的存活率与表达基因中的CPD修复之间存在相关性。因此,我们证明了总体修复测量可能会产生误导。我们的结果对确定CPD和6-4PP的相对生物学重要性具有启示意义,并且进一步支持了从转录活性DNA中去除CPD对紫外线抗性至关重要的假设。