Smerdon M J, Lieberman M W
Biochemistry. 1980 Jun 24;19(13):2992-3000. doi: 10.1021/bi00554a025.
We have compared the initial distribution and subsequent redistribution within chromatin of nucleotides incorporated during the early ("rapid") phase and the late ("slow") phase of UV-induced DNA repair synthesis. As has been observed for the early repair phase, most or all of the nucleotides incorporated during the late repair phase are initially staphylococcal nuclease and DNase I "sensitive" (i.e., rapidly digested). This initial enhanced sensitivity is accompanied by both an underrepresentation of these nucleotides in the 145--165 base pari (core) DNA produced by staphylococcal nuclease digestion and an absence of these nucleotides in the approximately 20-base repeat pattern produced by DNase I digestion. Furthermore, nucleotides incorporated at late time after damage are involved in nucleosome rearrangements as reported previously for repair synthesis occurring at early times [Smerdon, M. J., & Lieberman, M. W. (1978) Proc. Natl. Acad. Sci. U.S.A. 75, 4238--4241]. The kinetics of redistribution, however, appear to be more rapid than those observed for early times. Following redistribution the average nucleosome repeat length of DNA containing repair-incorporated nucleotides is the same as that of bulk DNA regardless of the time after damage that repair occurs; also, many of these nucleotides coelectrophorese with the approximately 10-base repeat fragments generated by DNase I. The results yield a new interpretation of our previous studies [Smerdeon, M. J., Tlsty, T. D., & Lieberman, M. W. (1978) Biochemistry 17, 2377--2386] on the distribution of nucleotides incorporated at long times after UV irradiation.
我们比较了紫外线诱导的DNA修复合成早期(“快速”)阶段和晚期(“缓慢”)阶段掺入染色质中的核苷酸的初始分布及其随后的重新分布。正如早期修复阶段所观察到的那样,晚期修复阶段掺入的大多数或所有核苷酸最初对葡萄球菌核酸酶和DNase I“敏感”(即快速消化)。这种初始增强的敏感性伴随着这些核苷酸在葡萄球菌核酸酶消化产生的145 - 165碱基对(核心)DNA中含量不足,以及在DNase I消化产生的约20碱基重复模式中缺乏这些核苷酸。此外,损伤后晚期掺入的核苷酸参与核小体重排,这与之前报道的早期发生的修复合成情况相同[Smerdon, M. J., & Lieberman, M. W. (1978) Proc. Natl. Acad. Sci. U.S.A. 75, 4238 - 4241]。然而,重新分布的动力学似乎比早期观察到的更快。重新分布后,含有修复掺入核苷酸的DNA的平均核小体重复长度与总体DNA相同,无论修复发生在损伤后的何时;而且,许多这些核苷酸与DNase I产生的约10碱基重复片段共电泳。这些结果对我们之前关于紫外线照射后长时间掺入的核苷酸分布的研究[Smerdeon, M. J., Tlsty, T. D., & Lieberman, M. W. (1978) Biochemistry 17, 2377 - 2386]产生了新的解释。