一种用于分析前期细胞核解体的分级无细胞体系。
A fractionated cell-free system for analysis of prophase nuclear disassembly.
作者信息
Suprynowicz F A, Gerace L
出版信息
J Cell Biol. 1986 Dec;103(6 Pt 1):2073-81. doi: 10.1083/jcb.103.6.2073.
Abstract
We describe a cell-free system in which a postribosomal supernatant (s140) from metaphase Chinese hamster ovary (CHO) cells induces prophase-like changes in isolated CHO cell nuclei, including chromatin condensation, and nuclear envelope and lamina disassembly. These events are strongly promoted by gamma-S-ATP and an ATP-regenerating system, and do not take place with an s140 derived from G2-phase cells. The metaphase cell s140 also induces disassembly of an isolated nuclear lamina fraction that is depleted of membranes, chromatin, and nuclear pore complexes. Disassembly of the isolated lamina is accompanied by phosphorylation of the major lamina proteins (lamins A, B, and C) to levels characteristic of metaphase cells. Kinetic analysis of lamina depolymerization indicates that cooperativity may be involved in this process. The biochemical properties of in vitro lamina disassembly suggest that the activity that depolymerizes the lamina during mitosis is soluble in metaphase cells, and support the notion that this activity is a lamin protein kinase.
摘要
我们描述了一种无细胞体系，其中从中期中国仓鼠卵巢（CHO）细胞获得的核糖体后上清液（s140）可诱导分离出的CHO细胞核发生类似前期的变化，包括染色质凝聚以及核膜和核纤层解体。γ-S-ATP和ATP再生系统可强烈促进这些事件的发生，而源自G2期细胞的s140则不会引发这些变化。中期细胞的s140还可诱导分离出的不含膜、染色质和核孔复合体的核纤层组分解体。分离出的核纤层解体伴随着主要核纤层蛋白（核纤层蛋白A、B和C）磷酸化至中期细胞特有的水平。核纤层解聚的动力学分析表明，协同作用可能参与了这一过程。体外核纤层解体的生化特性表明，在有丝分裂期间使核纤层解聚的活性物质在中期细胞中是可溶的，这支持了该活性物质是一种核纤层蛋白激酶的观点。
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