Delort A M, Duplaa A M, Molko D, Teoule R, Leblanc J P, Laval J
Nucleic Acids Res. 1985 Jan 25;13(2):319-35. doi: 10.1093/nar/13.2.319.
Various octadeoxynucleotides containing uracil at different positions were synthesized and submitted to the action of Escherichia coli and Micrococcus luteus uracil-DNA glycosylases. A uracil residue situated at the 5'-end was excised by the M.luteus enzyme but not by the E.coli one. Uracil residues located at the ultimate and penultimate positions at the 3'-end were not cleaved by either enzymes. At the other central positions, uracil was eliminated with different initial velocities. Single stranded phi X 174 DNA fragments were used to study the influence of the sequence. Cytosine bases were deaminated to give uracil by bisulfite treatment. It was shown that the initial excision velocity of two vicinal uracil residues was decreased. The same observation was made for two uracils separated by one base. A hypothetical scheme is suggested to explain the mechanism of action of uracil-DNA glycosylases.
合成了在不同位置含有尿嘧啶的各种十八聚脱氧核苷酸，并将其置于大肠杆菌和藤黄微球菌尿嘧啶-DNA糖基化酶的作用下。位于5'-末端的尿嘧啶残基被藤黄微球菌酶切除，但不被大肠杆菌酶切除。位于3'-末端最后和倒数第二个位置的尿嘧啶残基均未被这两种酶切割。在其他中心位置，尿嘧啶以不同的初始速度被去除。使用单链φX 174 DNA片段来研究序列的影响。通过亚硫酸氢盐处理将胞嘧啶碱基脱氨生成尿嘧啶。结果表明，两个相邻尿嘧啶残基的初始切除速度降低。对于被一个碱基隔开的两个尿嘧啶也有相同的观察结果。提出了一个假设方案来解释尿嘧啶-DNA糖基化酶的作用机制。
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