核糖体基因在分离的核小体核心颗粒中的脱氧核糖核酸酶I敏感性
DNase I sensitivity of ribosomal genes in isolated nucleosome core particles.
作者信息
Giri C P, Gorovsky M A
出版信息
Nucleic Acids Res. 1980 Jan 11;8(1):197-214. doi: 10.1093/nar/8.1.197-e.
Abstract
The level of chromatin structure at which DNase I recognizes conformational differences between inert and activated genes has been investigated. Bulk and ribosomal DNA's of Tetrahymena pyriformis were differentially labeled in vivo with [14C]- and [3H]-thymidine, respectively, utilizing a defined starvation-refeeding protocol. The 3H-labeled ribosomal genes were shown to be preferentially digested by DNase I in isolated nuclei. Staphylococcal nuclease digested the ribosomal genes more slowly than bulk DNA, probably owing to the higher GC content of rDNA. DNase I and staphylococcal nuclease digestions of purified nucleosomes and of nucleosome core particles isolated from dual-labeled, starved-refed nuclei were indistinguishable from those of intact nuclei. We conclude from these studies that DNase I recognizes an alteration in the internal nucleosome core structure of activated ribosomal genes.
摘要
已经对脱氧核糖核酸酶I识别惰性基因和活化基因之间构象差异的染色质结构水平进行了研究。利用确定的饥饿-再喂养方案，分别用[14C]-和[3H]-胸苷在体内对梨形四膜虫的总DNA和核糖体DNA进行差异标记。结果表明，在分离的细胞核中，3H标记的核糖体基因优先被脱氧核糖核酸酶I消化。葡萄球菌核酸酶消化核糖体基因的速度比总DNA慢，这可能是由于核糖体DNA的GC含量较高。从双标记、饥饿-再喂养的细胞核中分离出的纯化核小体和核小体核心颗粒的脱氧核糖核酸酶I和葡萄球菌核酸酶消化情况与完整细胞核的情况没有区别。我们从这些研究中得出结论，脱氧核糖核酸酶I识别活化核糖体基因内部核小体核心结构的改变。
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