Swiss Institute for Experimental Cancer Research (ISREC), Chemin des Boveresses 155, CH-1066 Epalinges/Lausanne, Switzerland.
J Cell Sci. 1990 Jul;96(Pt 3):439-50. doi: 10.1242/jcs.96.3.439.
The yeast nuclear scaffold has been shown to bind with high affinity to genomic sequences that permit autonomous DNA replication of plasmids (ARS elements). We describe here conditions for the isolation of a histone-free nuclear substructure, the nuclear scaffold, from Saccharomyces cerevisiae. We examine the protein composition of this structure,and the conditions under which topoisomerase II, the nuclear factor RAP-1 and RNA polymerase II co-fractionate with the scaffold. We find that exposure of nuclei to a combined metal and heat treatment (0.5mM Cu(2) +, 37 degree centigrade prior to detergent extraction is required for effective stabilization of these proteins with the scaffold. Electron microscopy of the residual nuclei extracted with non-ionic detergents shows the absence of a typical peripheral lamina structure.
酵母核骨架已被证明能与高亲和力结合，使质粒（ARS 元件）的自主 DNA 复制的基因组序列。我们在这里描述了从酿酒酵母中分离无组蛋白核亚结构核骨架的条件。我们检查了这种结构的蛋白质组成，以及拓扑异构酶 II、核因子 RAP-1 和 RNA 聚合酶 II 与支架共分离的条件。我们发现，核暴露于联合金属和热处理（0.5mM Cu(2) +，37 摄氏度）在去污剂提取之前是稳定这些蛋白质与支架的必要条件。用非离子型去污剂提取的残余核的电子显微镜观察显示，典型的外周层结构不存在。
