Caplan A, Ord M G, Stocken L A
Biochem J. 1978 Aug 15;174(2):475-83. doi: 10.1042/bj1740475.
Liver nuclei were prepared through the first cell cycle in partially hepatectomized young rats showing 30% parenchymal cell synchrony. To determine if nucleosome structure altered during this period, liver nuclei from sham-operated rats were compared with nuclei isolated at various times after partial hepatectomy. These nuclei were exposed to deoxyribonuclease I (EC 3.1.4.5), deoxyribonuclease II (EC 3.1.4.6) or micrococcal nuclease (EC 3.1.4.7) and the nucleosome-associated DNA length was ascertained. In no case was a difference in the DNA lengths associated with nucleosome structure observed. Differences were observed with regard to the histones and their relative association with nuclear material. When nuclei from normal rat livers were incubated in hypo-osmolar medium 9% of histone 1 and 4% of the other histones were released. These released histones, unlike those remaining bound to the nuclei, showed high [3H]adenosine and [3H]acetate uptakes in vivo. [32P]P1 uptake was also much greater into released than bound histones 1 and 3, but was not different for histone2A. At 3.5-4.5 h after partial hepatectomy, the release of histone 1 was trebled and that of histone 4 doubled. By 13.5 h, when phosphorylation of the bound forms of histones 2A and especially 1 was increased, no further changes in histone release in hypo-osmolar medium were found. The released histones from partially hepatectomized livers had indistinguishable [3H]adenosine uptakes from controls. The roles are discussed of phosphorylation and ADP-ribosylation in labilizing histone binding.
在部分肝切除的幼鼠中制备肝细胞核,这些幼鼠的实质细胞同步率为30%,处于第一个细胞周期。为了确定在此期间核小体结构是否发生改变,将假手术大鼠的肝细胞核与部分肝切除后不同时间分离的细胞核进行比较。将这些细胞核暴露于脱氧核糖核酸酶I(EC 3.1.4.5)、脱氧核糖核酸酶II(EC 3.1.4.6)或微球菌核酸酶(EC 3.1.4.7),并确定与核小体相关的DNA长度。在任何情况下,均未观察到与核小体结构相关的DNA长度存在差异。在组蛋白及其与核物质的相对结合方面观察到了差异。当正常大鼠肝脏的细胞核在低渗培养基中孵育时,9%的组蛋白1和4%的其他组蛋白被释放。这些释放的组蛋白与那些仍与细胞核结合的组蛋白不同,在体内显示出较高的[3H]腺苷和[3H]乙酸摄取。释放的组蛋白1和3对[32P]P1的摄取也比结合的组蛋白大得多,但组蛋白2A则没有差异。部分肝切除后3.5 - 4.5小时,组蛋白1的释放增加了两倍,组蛋白4的释放增加了一倍。到13.5小时,当组蛋白2A尤其是组蛋白1的结合形式的磷酸化增加时,在低渗培养基中组蛋白释放没有进一步变化。部分肝切除肝脏释放的组蛋白与对照组的[3H]腺苷摄取没有区别。讨论了磷酸化和ADP核糖基化在使组蛋白结合不稳定中的作用。