Glover T W, Chang C C, Trosko J E, Li S S
Proc Natl Acad Sci U S A. 1979 Aug;76(8):3982-6. doi: 10.1073/pnas.76.8.3982.
The UV induction of diphtheria toxin-resistant (DTr) mutants in normal and xeroderma pigmentosum human fibroblasts has been quantitatively characterized. A concentration of diphtheria toxin at which DTr cells are cross-resistant to Pseudomonas aeruginosa exotoxin A was determined and used in the selection of resistant mutants. Recovery of mutants was not influenced by the presence of wild-type cell densities of 1-8 x 10(5) per 9-cm plate, indicating no metabolic cooperation exists, in contrast to what is seen in the selection of some other variant phenotypes. Expression periods for UV-induced mutations differed with the severity of mutagen treatment and cell strain used. A relatively long (10-15 days after UV treatment) expression period was required for the maximum recovery of DTr mutants. Maximum recovery was followed by a decrease in mutation frequency on subsequent days evaluated. An apparent linear dose response within the dose range used was observed for UV-induced mutations in both normal and xeroderma pigmentosum fibroblasts. Our results indicate that xeroderma pigmentosum fibroblasts have higher UV-induced mutation frequencies per unit UV dose but similar frequencies per unit survival compared to normal cells within the range of UV doses tested.
已对正常和着色性干皮病人类成纤维细胞中抗白喉毒素(DTr)突变体的紫外线诱导进行了定量表征。确定了DTr细胞对铜绿假单胞菌外毒素A具有交叉抗性的白喉毒素浓度,并用于选择抗性突变体。每9厘米培养皿中1-8×10⁵个野生型细胞密度的存在并不影响突变体的恢复,这表明不存在代谢合作,这与在选择其他一些变异表型时所观察到的情况相反。紫外线诱导突变的表达期因诱变处理的严重程度和所用细胞株而异。DTr突变体的最大恢复需要相对较长的表达期(紫外线处理后10-15天)。最大恢复之后,在随后评估的几天中突变频率下降。在所用剂量范围内,正常和着色性干皮病成纤维细胞中紫外线诱导的突变均呈现明显的线性剂量反应。我们的结果表明,在测试的紫外线剂量范围内,与正常细胞相比,着色性干皮病成纤维细胞每单位紫外线剂量的紫外线诱导突变频率更高,但每单位存活率的频率相似。