Yagi T, Matsumura Y, Sato M, Nishigori C, Mori T, Sijbers A M, Takebe H
Department of Radiation Genetics, Graduate School of Medicine, Kyoto University, Japan.
Carcinogenesis. 1998 Jan;19(1):55-60. doi: 10.1093/carcin/19.1.55.
XP-F cDNA was cloned into a mammalian expression vector plasmid, and introduced into group F xeroderma pigmentosum (XP-F) cells. Several cell clones possessing transfected XPF cDNA were randomly isolated, and DNA repair characteristics of a clone, XP-FR2, were extensively analyzed. The XP-FR2 cells expressed high level of XPF protein as well as ERCC1 protein, although their parental XP-F cells expressed extremely low level of both proteins. The XP-FR2 cells showed UV resistance comparable to normal human cells, and had normal levels of UV-induced unscheduled DNA synthesis and normal capability to remove cyclobutane pyrimidine dimers and (6-4) photoproducts. Frequencies and types of UV-induced mutations examined by shuttle vector plasmids in XP-FR2 cells were similar to those in normal human cells. These results demonstrate that excision repair defect in XP-F cells is fully corrected by over-expression of XPF cDNA alone, although only partial correction of the cells by XPF cDNA has been reported before.
将XP-F cDNA克隆到哺乳动物表达载体质粒中,并导入F组着色性干皮病(XP-F)细胞。随机分离出几个含有转染XPF cDNA的细胞克隆,并对其中一个克隆XP-FR2的DNA修复特性进行了广泛分析。XP-FR2细胞表达高水平的XPF蛋白以及ERCC1蛋白,尽管其亲本XP-F细胞中这两种蛋白的表达水平极低。XP-FR2细胞表现出与正常人细胞相当的紫外线抗性,紫外线诱导的非定标DNA合成水平正常,去除环丁烷嘧啶二聚体和(6-4)光产物的能力也正常。通过穿梭载体质粒检测的XP-FR2细胞中紫外线诱导突变的频率和类型与正常人细胞相似。这些结果表明,仅通过XPF cDNA的过表达就可以完全纠正XP-F细胞中的切除修复缺陷,尽管之前报道过XPF cDNA只能部分纠正这些细胞。