IMMUNOCYTOCHEMICAL LOCALIZATION OF MAJOR POLYPEPTIDES OF NUCLEAR-PORE COMPLEX-LAMINA FRACTION - INTERPHASE AND MITOTIC DISTRIBUTION
IMMUNOCYTOCHEMICAL LOCALIZATION OF MAJOR POLYPEPTIDES OF NUCLEAR-PORE COMPLEX-LAMINA FRACTION - INTERPHASE AND MITOTIC DISTRIBUTION
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DOI：
10.1083/jcb.79.2.546
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发表时间：
1978-01-01
影响因子：
7.8
通讯作者：
BLOBEL, G
中科院分区：
文献类型：
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作者：
GERACE, L;BLUM, A;BLOBEL, G
以前从大鼠肝细胞核中分离出一种组分，该组分由与内核膜下的蛋白质层相关的核孔复合体组成。利用十二烷基硫酸钠(十二烷基硫酸钠)凝胶洗脱出的蛋白质，在鸡体内制备了针对3条主要毛孔复合层带的抗体。Ouchterlony双扩散分析表明，这些单条带中的每一条都与3种抗血清发生强烈的交叉反应。在用这些抗体对组织培养细胞进行免疫荧光定位时，在间期核的周围获得了强烈的染色模式，几乎没有细胞质反应。大鼠肝细胞核的EM免疫过氧化物酶染色，这些抗体仅标记核周。反应发生在含有板层的区域，但不发生在孔道复合体。虽然分离程序完全提取了细胞核的内部内容物，但半定量的Ouchterlony分析表明，它释放的这些板层抗原的量可以忽略不计。这3条带代表了外周核板的主要成分，而不是内部核蛋白基质的结构元素。荧光显微镜显示，在有丝分裂过程中，这些板层蛋白的核周间期定位发生了巨大的变化。伴随着早期核膜的解体，这些抗原在整个细胞内呈弥漫性定位。这种分布一直持续到末期，此时抗原逐渐并完全定位在子代染色体团的表面。椎板是一种生物聚合物，在有丝分裂过程中可以进行可逆分解。
A fraction from rat liver nuclei consisting of nuclear pore complexes associated with the proteinaceous lamina which underlies the inner nuclear membrane was previously isolated. Using protein eluted from sodium dodecyl sulfate (SDS) gels, antibodies in chickens were prepared to each of the 3 predominant pore complex-lamina bands. Ouchterlony double diffusion analysis shows that each of these individual bands cross-reacts strongly with all 3 antisera. In immunofluorescence localization performed on tissue culture cells with these antibodies a pattern of intense staining at the periphery of the interphase nucleus was obtained, with little or no cytoplasmic reaction. EM immunoperoxidase staining of rat liver nuclei with these antibodies labeled the nuclear periphery exclusively. Reaction occurred in areas which contain the lamina, but not at the pore complexes. While the isolation procedure extracts the internal contents of nuclei completely, semiquantitative Ouchterlony analysis showed that it released negligible amounts of these lamina antigens. These 3 bands represented major components of a peripheral nuclear lamina, and were not structural elements of an internal nuclear protein matrix. Fluorescence microscopy showed that the perinuclear interphase localization of these lamina proteins undergoes dramatic changes during mitosis. Concomitant with nuclear envelope disassembly in prophase, these antigens assumed a diffuse localization throughout the cell. This distribution persists until telophase, when the antigens became progressively and completely localized at the surface of the daughter chromosome masses. The lamina is a biological polymer which can undergo reversible disassembly during mitosis.