Nuclear events of apoptosis in vitro in cell-free mitotic extracts: a model system for analysis of the active phase of apoptosis.
Nuclear events of apoptosis in vitro in cell-free mitotic extracts: a model system for analysis of the active phase of apoptosis.
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DOI：
10.1083/jcb.123.1.7
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发表时间：
1993-10
影响因子：
7.8
通讯作者：
Earnshaw, W C
中科院分区：
文献类型：
--
作者：
Lazebnik, Y A;Cole, S;Cooke, C A;Nelson, W G;Earnshaw, W C
我们已经开发了一个无细胞系统，诱导的形态学转变的特点，在孤立的细胞核凋亡。该系统使用从有丝分裂的鸡肝癌细胞中制备的提取物，随后进行连续的S期/M期同步化。当将细胞核加入这些提取物中时，染色质高度浓缩成球形结构域，最终挤出核膜，形成凋亡小体。该过程是高度同步的，结构变化在60分钟内完成。与这些形态学变化相一致，核DNA被切割成核小体梯。这两个过程都受到抑制的Zn 2+，在完整的细胞凋亡的抑制剂。核纤层拆卸伴随着这些结构的变化，在添加的核，我们表明，纤层拆卸是一个典型的功能在小鼠，人类和鸡的完整细胞凋亡。这个系统可能提供一个强大的手段解剖的生化机制的最后阶段的细胞凋亡。
We have developed a cell-free system that induces the morphological transformations characteristic of apoptosis in isolated nuclei. The system uses extracts prepared from mitotic chicken hepatoma cells following a sequential S phase/M phase synchronization. When nuclei are added to these extracts, the chromatin becomes highly condensed into spherical domains that ultimately extrude through the nuclear envelope, forming apoptotic bodies. The process is highly synchronous, and the structural changes are completed within 60 min. Coincident with these morphological changes, the nuclear DNA is cleaved into a nucleosomal ladder. Both processes are inhibited by Zn2+, an inhibitor of apoptosis in intact cells. Nuclear lamina disassembly accompanies these structural changes in added nuclei, and we show that lamina disassembly is a characteristic feature of apoptosis in intact cells of mouse, human and chicken. This system may provide a powerful means of dissecting the biochemical mechanisms underlying the final stages of apoptosis.