van Duin M, Westerveld A, Hoeijmakers J H
Mol Cell Biol. 1985 Apr;5(4):734-41. doi: 10.1128/mcb.5.4.734-741.1985.
Irradiation of dominant marker DNA with UV light (150 to 1,000 J/m2) was found to stimulate the transformation of human cells by this marker from two- to more than fourfold. This phenomenon is also displayed by xeroderma pigmentosum cells (complementation groups A and F), which are deficient in the excision repair of UV-induced pyrimidine dimers in the DNA. Also, exposure to UV of the transfected (xeroderma pigmentosum) cells enhanced the transfection efficiency. Removal of the pyrimidine dimers from the DNA by photoreactivating enzyme before transfection completely abolished the stimulatory effect, indicating that dimer lesions are mainly responsible for the observed enhancement. A similar stimulation of the transformation efficiency is exerted by 2-acetoxy-2-acetylaminofluorene modification of the DNA. No stimulation was found after damaging vector DNA by treatment with DNase or gamma rays. These findings suggest that lesions which are targets for the excision repair pathway induce the increase in transformation frequency. The stimulation was found to be independent of sequence homology between the irradiated DNA and the host chromosomal DNA. Therefore, the increase of the transformation frequency is not caused by a mechanism inducing homologous recombination between these two DNAs. UV treatment of DNA before transfection did not have a significant effect on the amount of DNA integrated into the xeroderma pigmentosum genome.
发现用紫外线(150至1000 J/m²)照射显性标记DNA可使该标记诱导的人类细胞转化增加两倍至四倍以上。着色性干皮病细胞(互补组A和F)也表现出这种现象,这些细胞在DNA中紫外线诱导的嘧啶二聚体的切除修复方面存在缺陷。此外,转染的(着色性干皮病)细胞暴露于紫外线也提高了转染效率。在转染前用光复活酶去除DNA中的嘧啶二聚体完全消除了刺激作用,表明二聚体损伤是观察到的增强作用的主要原因。DNA经2-乙酰氧基-2-乙酰氨基芴修饰也对转化效率有类似的刺激作用。用DNase或γ射线处理破坏载体DNA后未发现刺激作用。这些发现表明,作为切除修复途径靶点的损伤会导致转化频率增加。发现这种刺激与照射的DNA和宿主染色体DNA之间的序列同源性无关。因此,转化频率的增加不是由诱导这两种DNA之间同源重组的机制引起的。转染前对DNA进行紫外线处理对整合到着色性干皮病基因组中的DNA量没有显著影响。