Use of exonuclease III to determine the site of stable lesions in defined sequences of DNA: the cyclobutane pyrimidine dimer and cis and trans dichlorodiammine platinum II examples.
Use of exonuclease III to determine the site of stable lesions in defined sequences of DNA: the cyclobutane pyrimidine dimer and cis and trans dichlorodiammine platinum II examples.
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使用核酸外切酶 III 确定 DNA 确定序列中稳定损伤的位点：环丁烷嘧啶二聚体以及顺式和反式二氯二氨铂 II 示例。
DOI：
10.1093/nar/9.18.4595
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发表时间：
1981
影响因子：
14.9
通讯作者：
Haseltine,WA
中科院分区：
文献类型：
--
作者：
Royer-Pokora,B;Gordon,LK;Haseltine,WA
利用双链特异性核酸内切酶Ⅲ消化5′-末端标记的DNA，建立了一种检测DNA中化学稳定性损伤的方法。当在高分辨率DNA测序凝胶上分析时，产物揭示了DNA修饰的位点。通过比较外切核酸酶III消化产生的片段与用UV特异性内切核酸酶消化DNA后获得的片段，可以定位由UV照射诱导的环丁烷嘧啶二聚体。实验证明，核酸外切酶III在离环丁烷嘧啶二聚体一个碱基处终止。用顺式和反式二氯二氨铂（II）进行的类似实验表明，这些试剂对DNA的修饰也会阻碍核酸外切酶III的消化。一般来说，顺式和反式铂加合物的终止位点相同。它们出现在鸟嘌呤碱基的位点。在相邻鸟嘌呤碱基的位点发现了其他的终止位点。这些结果与顺铂形成链内鸟嘌呤-鸟嘌呤二聚体的模型一致，而反式铂不形成。
A method to detect chemically stable lesions in DNA has been developed using Exonuclease III, a double strand specific nuclease, to digest 5′-end labeled DNA. The products, when analyzed on high resolution DNA sequencing gels, reveal the sites of DNA modification. Cyclobutane pyrimidine dimers induced by UV irradiation can be localized by comparison of the fragments produced by Exonuclease III digestion with fragments obtained after digestion of the DNA with UV specific endonuclease. The experiments demonstrate that Exonuclease III stops one base away from the cyclobutane pyrimidine dimers. Similar experiments withcis- andtrans-dichlorodiammine-platinum (II) showed that modifications of DNA by these agents also impede Exonuclease III digestion. In general the same stop sites were found forcis-andtrans-platinum adducts. They occur at sites of guanine bases. Additional stop sites were found forcis-platinum at sites of adjacent guanine bases. These results are in agreement with the model thatcis-platinum forms intrastrand guanine-guanine dimers, whereastrans-platinum does not.