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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Acknowledgements
This work was supported by JSPS KAKENHI and MEXT KAKENHI, Grant in Aid from the Ministry of Education, Culture, Sports, Science, and Technology of Japan.
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Nishitani, H., Morino, M., Murakami, Y., Maeda, T., Shiomi, Y. (2014). Chromatin Fractionation Analysis of Licensing Factors in Mammalian Cells. In: Noguchi, E., Gadaleta, M. (eds) Cell Cycle Control. Methods in Molecular Biology, vol 1170. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0888-2_28
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DOI: https://doi.org/10.1007/978-1-4939-0888-2_28
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