Chapter title |
Chromatin Fractionation Analysis of Licensing Factors in Mammalian Cells.
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Chapter number | 28 |
Book title |
Cell Cycle Control
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Published in |
Methods in molecular biology, June 2014
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DOI | 10.1007/978-1-4939-0888-2_28 |
Pubmed ID | |
Book ISBNs |
978-1-4939-0887-5, 978-1-4939-0888-2
|
Authors |
Nishitani H, Morino M, Murakami Y, Maeda T, Shiomi Y, Hideo Nishitani, Masayuki Morino, Yusuke Murakami, Takeshi Maeda, Yasushi Shiomi |
Abstract |
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. |
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