Bailleul B, Galiègue-Zouitina S, Loucheux-Lefebvre M H
Nucleic Acids Res. 1984 Oct 25;12(20):7915-27. doi: 10.1093/nar/12.20.7915.
Poly(dG-dC).poly(dG-dC) has been modified by reaction with 4-acetoxyaminoquinoline 1-oxide (Ac-4 HAQO), the ultimate carcinogen of 4-nitroquinoline 1-oxide. The circular dichroism (CD) spectra of the modified and unmodified polymers have been compared under various experimental conditions. The CD spectra were recorded in 1 mM phosphate, 50% (v/v) ethanol, 3.8 M LiCl and 95% (v/v) ethanol, conditions in which poly(dG-dC).poly(dG-dC) adopts the B-, Z-, C- and A-form respectively. In 1 mM phosphate buffer, poly(dG-dC).poly(dG-dC) modified by Ac-4 HAQO seems not to contain regions in the Z-form. Z-form induction could be progressively obtained by the addition of ethanol as follows: in the buffer with about 30% ethanol the modified polymer started to adopt the Z structure, while 40% of ethanol in the buffer was necessary for the unmodified polymer. In the 50% ethanol-1 mM phosphate buffer mixture (v/v), poly(dG-dC).poly(dG-dC) was entirely in the Z-form while poly(dG-dC).poly(dG-dC) modified by Ac-4 HAQO remained partially in the B-form. Enzymatic digestions with the nuclease S1 which is specific of the single-stranded DNA were carried out in order to support the modified poly(dG-dC).poly(dG-dC) CD study conclusions. The role played by the two major adducts on the conformational characteristics of modified polymer is discussed.
聚(dG-dC)·聚(dG-dC)已通过与4-乙酰氧基氨基喹啉1-氧化物(Ac-4 HAQO)反应进行了修饰,Ac-4 HAQO是4-硝基喹啉1-氧化物的最终致癌物。在各种实验条件下比较了修饰和未修饰聚合物的圆二色性(CD)光谱。CD光谱是在1 mM磷酸盐、50%(v/v)乙醇、3.8 M LiCl和95%(v/v)乙醇中记录的,在这些条件下聚(dG-dC)·聚(dG-dC)分别呈现B型、Z型、C型和A型。在1 mM磷酸盐缓冲液中,经Ac-4 HAQO修饰的聚(dG-dC)·聚(dG-dC)似乎不包含Z型区域。通过添加乙醇可以逐步诱导Z型形成,具体如下:在含有约30%乙醇的缓冲液中,修饰后的聚合物开始呈现Z结构,而未修饰的聚合物则需要缓冲液中40%的乙醇。在50%乙醇-1 mM磷酸盐缓冲液混合物(v/v)中,聚(dG-dC)·聚(dG-dC)完全呈Z型,而经Ac-4 HAQO修饰的聚(dG-dC)·聚(dG-dC)仍部分呈B型。用对单链DNA具有特异性的核酸酶S1进行酶切消化,以支持对修饰后的聚(dG-dC)·聚(dG-dC)的CD研究结论。讨论了两种主要加合物对修饰后聚合物构象特征所起的作用。