Chromatin fractionation analysis of licensing factors in mammalian cells.
Chromatin fractionation analysis of licensing factors in mammalian cells.
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DOI：
10.1007/978-1-4939-0888-2_28
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发表时间：
2014
影响因子：
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通讯作者：
H. Nishitani;Masayuki Morino;Y. Murakami;Takeshi Maeda;Y. Shiomi
中科院分区：
文献类型：
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作者：
H. Nishitani;Masayuki Morino;Y. Murakami;Takeshi Maeda;Y. Shiomi
ORC、Cdc 6、Cdt 1和MCM 2 -7是复制许可因子，它们在每细胞周期一次的DNA复制控制中发挥核心作用。ORC、Cdc 6和Cdt 1协作将MCM 2 -7加载到复制起点，以便许可它们进行复制。MCM 2 -7是直接参与DNA复制的DNA解旋酶，并且随着S期的进展和每个复制子的复制而从DNA解离。在细胞周期中，MCM 2 -7的负载在有丝分裂结束和G1期受到限制。因此，染色质结合的MCM 2 -7及其负载物的水平在细胞周期期间振荡。这些因子的染色质缔合可以通过将细胞裂解物分离成哺乳动物细胞中的可溶性和富含染色质的不溶性级分来分析。
ORC, Cdc6, Cdt1, and MCM2-7 are replication-licensing factors, which play a central role in the once-per-cell cycle control of DNA replication. ORC, Cdc6, and Cdt1 collaborate to load MCM2-7 onto replication origins in order to license them for replication. MCM2-7 is a DNA helicase directly involved in DNA replication and dissociates from DNA as S phase progresses and each replicon is replicated. In the cell cycle, the loading of MCM2-7 is restricted during the end of mitosis and the G1 phase. Thus, the levels of chromatin-bound MCM2-7 and its loaders oscillate during the cell cycle. Chromatin association of these factors can be analyzed by separating a cell lysate into soluble and chromatin-enriched insoluble fractions in mammalian cells.