Varshavsky A J, Georgiev G P
Mol Biol Rep. 1976 Sep;3(1):27-38. doi: 10.1007/BF00357206.
Oligomers of chromatin subunits (oligonucleosomes) were prepared by a mild digestion of chromatin with staphylococcal nuclease followed by a purification of a high molecular weight material (hexanucleosomes and larger DNP particles) by gel chromatography. The main finding is that a mild removal of histone H1 from the oligonucleosome preparation by treatment with tRNA in the absence of any significant hydrodynamic shearing leads to the formation of free DNA molecules which constitute 5-6% of the total oligonucleosomal DNA. The size of nucleosome-free DNA stretches in H1-depleted hydrodynamically sheared chromatin is about 6000 base pairs and their content is apparently 10-12% of the total DNA. These and related findings are discussed in terms of the previously proposed "asymmetric hairpin" model of DNA packing in chromatin [1-4]. Different kinds of the asymmetric hairpin are considered and ambiguities in interpretations of experimental data are pointed out.
通过用葡萄球菌核酸酶温和消化染色质,随后通过凝胶色谱法纯化高分子量物质(六核小体和更大的脱氧核糖核蛋白颗粒),制备染色质亚基的寡聚体(寡核小体)。主要发现是,在不存在任何明显流体动力学剪切的情况下,通过用tRNA处理从寡核小体制剂中温和去除组蛋白H1会导致形成游离DNA分子,其占寡核小体总DNA的5-6%。在H1缺失的流体动力学剪切染色质中,无核小体DNA片段的大小约为6000个碱基对,其含量显然占总DNA的10-12%。根据先前提出的染色质中DNA堆积的“不对称发夹”模型[1-4]对这些及相关发现进行了讨论。考虑了不同类型的不对称发夹,并指出了实验数据解释中的模糊性。