Weber J L, Cole R D
J Biol Chem. 1982 Oct 10;257(19):11784-90.
Some of the properties of chromatographically purified satellite chromatin are compared with those of unfractionated, control chromatin. Nucleosomes were present in the purified satellite chromatin as verified by digestion with micrococcal nuclease and DNase I and by electron microscopy. Average nucleosome DNA repeat lengths of 186 +/- 7 and 193 +/- 5 base pairs were obtained through micrococcal nuclease digestion of the purified satellite chromatin and control chromatin, respectively; nucleosome spacer lengths were equally heterogeneous for the two chromatin samples. The distribution of Eco RI-produced chromatin fragments of different size in the satellite chromatin was the same as that calculated assuming random cleavage at each Eco RI site, consistent with the notion that nucleosomes do not have specific locations on the 1.715 g ml-1 satellite DNA. The purified satellite chromatin contained little non-histone protein, but did contain all five histones and all detectable histone sequence variants. Amounts of the core histones were identical in the satellite chromatin and the control chromatin, but the amount of histone H1 was 30% less in the satellite chromatin than in the control. Although the molar ratios for the major sequence variants of both histones H3 and H2A differed between kidney and thymus, the ratios were the same for satellite and control chromatin isolated from a single tissue.
将色谱纯化的卫星染色质的一些特性与未分级的对照染色质的特性进行了比较。经微球菌核酸酶和DNase I消化以及电子显微镜验证,纯化的卫星染色质中存在核小体。通过对纯化的卫星染色质和对照染色质进行微球菌核酸酶消化,分别获得了平均核小体DNA重复长度为186±7和193±5个碱基对;两种染色质样品的核小体间隔长度同样具有异质性。卫星染色质中不同大小的Eco RI产生的染色质片段的分布与假设在每个Eco RI位点随机切割所计算的分布相同,这与核小体在1.715 g/ml卫星DNA上没有特定位置的观点一致。纯化的卫星染色质几乎不含有非组蛋白,但确实含有所有五种组蛋白以及所有可检测到的组蛋白序列变体。卫星染色质和对照染色质中核心组蛋白的含量相同,但卫星染色质中组蛋白H1的含量比对照染色质少30%。尽管肾和胸腺中组蛋白H3和H2A的主要序列变体的摩尔比不同,但从单个组织分离的卫星染色质和对照染色质的比例相同。