Gantt R, Jones G M, Stephens E V, Baeck A E, Sanford K K
Biochim Biophys Acta. 1979 Dec 17;565(2):231-40. doi: 10.1016/0005-2787(79)90201-6.
Cool-white fluorescent light induces crosslinks in DNA when proliferating cells are exposed at 37 degrees C for 20 h to 4.6 J/m2/s in culture medium supplemented with fetal bovine serum. Using the Kohn alkaline elution technique, we now find that: 1. Increased light intensity increases DNA crosslinks. 2. The crosslinking is medium-mediated. 3. Oxygen enhances the crosslinking. 4. The extent of crosslinking is decreased at high cell density. 5. The crosslinks can be removed by digestion with proteinase K (0.02 to 0.50 mg/ml). 6. Human cell lines including those derived from adult prostate, fetal lung (IMR-90) and mixed fetal tissues are susceptible to light-induced crosslinks. 7. Crosslinkage is not decreased by addition of catalase to the medium and the effective wavelength is probably between 450 nm and 490 nm. From these results we conclude that the mechanism of light-induced crosslinks differs from that of light-induced chromatid breaks and that the major lesion observed is protein-DNA cross-linkage rather than DNA strand breaks.
当增殖细胞在补充有胎牛血清的培养基中于37℃暴露于4.6 J/m²/s的冷白色荧光下20小时时,会诱导DNA交联。使用科恩碱性洗脱技术,我们现在发现:1. 光强度增加会增加DNA交联。2. 交联是由培养基介导的。3. 氧气会增强交联。4. 在高细胞密度下交联程度会降低。5. 交联可以通过用蛋白酶K(0.02至0.50 mg/ml)消化去除。6. 包括源自成人前列腺、胎儿肺(IMR-90)和混合胎儿组织的人类细胞系对光诱导的交联敏感。7. 向培养基中添加过氧化氢酶不会降低交联,有效波长可能在450 nm至490 nm之间。从这些结果我们得出结论,光诱导交联的机制与光诱导染色单体断裂的机制不同,观察到的主要损伤是蛋白质-DNA交联而不是DNA链断裂。