利用人类校正核酸内切酶（校正核酸酶）检测烷基化DNA中的不同类型损伤。
Detection of different types of damage in alkylated DNA by means of human corrective endonuclease (correndonuclease).
作者信息
Duker N J, Teebor G W
出版信息
Proc Natl Acad Sci U S A. 1976 Aug;73(8):2629-33. doi: 10.1073/pnas.73.8.2629.
Abstract
Corrective endonuclease (correndunclease) activity of HeLa cells was assayed with alkylated DNA. Double-stranded, covalently closed DNA from phage PM II was treated with methyl methanesulfonate, N-methyl-N-nitrosourea, beta-propiolactone, or diepoxybutane to introduce alkylated bases and alkali-labile sites into the DNA. The damaged DNA was incubated with an extract of HeLa cells that catalyzes the formation of breaks at apurinic sites in double-stranded DNA. Methylated DNA was broken at every alkali-labile site by the HeLa correndonuclease, which indicated that these sites are similar to the apurinic sites produced by heating at acid pH. DNA alkylated with beta-propiolactone or diepoxybutane containing the same number of alkali-labile sites was broken to a far lesser extent. This indicates the presence of a second type of alkali-labile damage that is correndonuclease-insensitive.
摘要
用烷基化DNA检测了HeLa细胞的校正核酸内切酶（correndunclease）活性。来自噬菌体PM II的双链、共价闭合DNA用甲磺酸甲酯、N-甲基-N-亚硝基脲、β-丙内酯或双环氧丁烷处理，以将烷基化碱基和碱不稳定位点引入DNA中。将受损DNA与HeLa细胞提取物一起孵育，该提取物催化双链DNA中脱嘌呤位点处的断裂形成。HeLa校正核酸内切酶在每个碱不稳定位点处使甲基化DNA断裂，这表明这些位点类似于在酸性pH下加热产生的脱嘌呤位点。用β-丙内酯或双环氧丁烷烷基化且含有相同数量碱不稳定位点的DNA断裂程度要小得多。这表明存在第二种对校正核酸内切酶不敏感的碱不稳定损伤类型。
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