Riley D E, Keller J M
J Cell Sci. 1978 Aug;32:249-68. doi: 10.1242/jcs.32.1.249.
Non-membranous HeLa cell nuclear ghosts, representing non-membranous nuclear envelope or 'skeletal' components, have been examined in whole-mount fashion by transmission electron microscopy. Major components of the ghosts include annuli with inner and outer diameters of 43 and 90 nm, respectively, which are consistent in dimensions with nuclear pore complexes. Also present are rod-like images (260 nm in length and 50 nm in width or diameter) representing either previously unobserved nuclear structures, or condensations of repeating functional units not otherwise observable. The annular and rod-like images were also observed when various steps in the ghost isolation procedure, such as the use of detergents, 0.5 M MgCl2 and polylysine attachment of the ghosts to electron-microscope grids, were circumvented. The annular and rod-like images are connected into linear and polygonal arrays by strands (15-30 nm in width) that are sensitive to DNase I and DNase II but resistant to nuclease S1. Thus, although the non-membranous ghosts from HeLa cells are composed primarily of protein, enzymic dissection indicates that their gross integrity is markedly dependent on double-stranded DNA. Nuclear ghosts prepared from a wide range of species including mammals, birds and plants, exhibited essentially the same components and organization.
已通过透射电子显微镜以整装方式检查了非膜性海拉细胞核膜空壳,其代表非膜性核被膜或“骨架”成分。核膜空壳的主要成分包括内径和外径分别为43纳米和90纳米的环,其尺寸与核孔复合体一致。还存在杆状图像(长度为260纳米,宽度或直径为50纳米),其代表要么是以前未观察到的核结构,要么是重复功能单元的凝聚物,否则无法观察到。当绕过核膜空壳分离程序中的各个步骤,例如使用去污剂、0.5 M氯化镁以及将核膜空壳附着到电子显微镜网格上的聚赖氨酸时,也观察到了环状和杆状图像。环状和杆状图像通过宽度为15 - 30纳米的链连接成线性和多边形阵列,这些链对脱氧核糖核酸酶I和脱氧核糖核酸酶II敏感,但对核酸酶S1有抗性。因此,尽管来自海拉细胞的非膜性核膜空壳主要由蛋白质组成,但酶解分析表明它们的整体完整性明显依赖于双链DNA。从包括哺乳动物、鸟类和植物在内的多种物种制备的核膜空壳表现出基本相同的成分和组织。