A NEW METHOD FOR THE ISOLATION OF REPLICATIVE CHROMATIN - SELECTIVE DEPOSITION OF HISTONE ON BOTH NEW AND OLD DNA
A NEW METHOD FOR THE ISOLATION OF REPLICATIVE CHROMATIN - SELECTIVE DEPOSITION OF HISTONE ON BOTH NEW AND OLD DNA
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DOI：
10.1016/0092-8674(81)90277-4
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发表时间：
1981-01-01
期刊：
影响因子：
64.5
通讯作者：
CHALKLEY, R
中科院分区：
文献类型：
--
作者：
JACKSON, V;CHALKLEY, R
建立了用甲醛固定全细胞（肝癌组织培养）后分离亚细胞成分的新方法。通过一些标准确定，固定不会改变或引起新复制或旧染色质的核小体结构重排。基于密度的差异，这种方法几乎可以完全从先前存在的物质中分离出新复制的染色质。新复制的染色质（甚至来自环己亚胺处理的细胞）似乎在两个子链上都含有核小体。通过利用分离新合成的染色质的能力，新的组蛋白沉积在复制叉的问题被重新审视。新合成的组蛋白H3和H4沉积在新的DNA上，并在那里停留很长时间。新H1沉积在旧DNA和新H2A-H2B上，虽然它们可能与新DNA短暂结合，但它们在很大程度上与先前存在的染色质有关。
A new method was developed for isolating subcellular components after fixation of whole cells [hepatoma tissue culture] with formaldehyde. By a number of criteria it was established that the fixation does not alter or cause rearrangement of nucleosomal structure of either newly replicated or old chromatin. This approach can be used to almost completely resolve newly replicated chromatin from preexisting material on the basis of the difference in density. Newly replicated chromatin (even from cycloheximide-treated cells) appears to contain nucleosomes on both daughter strands. By exploiting the ability to separate newly synthesized chromatin, the question of the deposition of new histone at the replication fork was reexamined. Newly synthesized histones H3 and H4 are deposited onto new DNA and stay in place for a significant time. New H1 is deposited an old DNA and new H2A-H2B, while they may be transiently bound to new DNA, are largely associated with preexisting chromatin.