Tsujimura T, Maher V M, Godwin A R, Liskay R M, McCormick J J
Department of Microbiology, Michigan State University, East Lansing 48824-1316.
Proc Natl Acad Sci U S A. 1990 Feb;87(4):1566-70. doi: 10.1073/pnas.87.4.1566.
To investigate the role of DNA damage and nucleotide excision repair in intrachromosomal homologous recombination, a plasmid containing duplicated copies of the gene coding for hygromycin resistance was introduced into the genome of a repair-proficient human cell line, KMST-6, and two repair-deficient lines, XP2OS(SV) from xeroderma pigmentosum complementation group A and XP2YO(SV) from complementation group F. Neither hygromycin-resistance gene codes for a functional enzyme because each contains an insertion/deletion mutation at a unique site, but recombination between the two defective genes can yield hygromycin-resistant cells. The rates of spontaneous recombination in normal and xeroderma pigmentosum cell strains containing the recombination substrate were found to be similar. The frequency of UV-induced recombination was determined for three of these cell strains. At low doses, the group A cell strain and the group F cell strain showed a significant increase in frequency of recombinants. The repair-proficient cell strain required 10- to 20-fold higher doses of UV to exhibit comparable increases in frequency of recombinants. These results suggest that unexcised DNA damage, rather than the excision repair process per se, stimulates such recombination.
为了研究DNA损伤和核苷酸切除修复在染色体内同源重组中的作用,将含有潮霉素抗性编码基因重复拷贝的质粒导入修复功能正常的人类细胞系KMST - 6以及两个修复缺陷细胞系,即来自A型着色性干皮病互补组的XP2OS(SV)和来自F型互补组的XP2YO(SV)的基因组中。两个潮霉素抗性基因均不编码功能性酶,因为每个基因在一个独特位点都含有插入/缺失突变,但两个缺陷基因之间的重组可产生潮霉素抗性细胞。含有重组底物的正常细胞系和着色性干皮病细胞系中的自发重组率相似。测定了其中三个细胞系紫外线诱导的重组频率。在低剂量时,A型细胞系和F型细胞系的重组体频率显著增加。修复功能正常的细胞系需要高10至20倍剂量的紫外线才能表现出可比的重组频率增加。这些结果表明,未切除的DNA损伤而非切除修复过程本身会刺激这种重组。