The nuclear matrix revealed by eluting chromatin from a cross-linked nucleus
The nuclear matrix revealed by eluting chromatin from a cross-linked nucleus
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DOI：
10.1073/pnas.94.9.4446
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发表时间：
1997-04-29
影响因子：
11.1
通讯作者：
Penman, S
中科院分区：
文献类型：
--
作者：
Nickerson, JA;Krockmalnic, G;Penman, S
细胞核是一个复杂的结构集成的许多功能域，其复杂的空间组织是由一个非染色质支架，核基质，我们在这里报告的方法制备的核基质与改进的超微结构的保存。在去除可溶性蛋白质后，核的结构与甲醛广泛交联。令人惊讶的是，染色质可以通过DNA酶I消化有效地去除，留下保存良好的核基质。通过该程序暴露的核基质由高度结构化的纤维组成，其连接到核纤层并建立在分支的10-nm核心细丝的底层网络上。染色质和核基质可以相对容易地分离，尽管先前进行了广泛的交联，这表明除了在染色质环的基部处的连接之外，两种结构之间几乎没有附着点，这是理解细胞核中染色质结构的重要线索。
The nucleus is an intricately structured integration of many functional domains whose complex spatial organization is maintained by a nonchromatin scaffolding, the nuclear matrix, We report here a method for preparing the nuclear matrix with improved preservation of ultrastructure. After the removal of soluble proteins, the structures of the nucleus were extensively cross-linked with formaldehylde. Surprisingly, the chromatin could be efficiently removed by DNase I digestion leaving a well preserved nuclear matrix. The nuclear matrix uncovered by this procedure consisted of highly structured fibers, connected to the nuclear lamina and built on an underlying network of branched IO-nm core filaments, The relative ease with which chromatin and the nuclear matrix could be separated despite extensive prior cross-linking suggests that there are few attachment points between the two structures other than the connections at the bases of chromatin loops, This is an important clue for understanding chromatin organization in the nucleus.