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Concerted Loading of Mcm2–7 Double Hexamers around DNA during DNA Replication Origin Licensing
Open accessAbstract
The licensing of eukaryotic DNA replication origins, which ensures once per cell cycle replication, involves the loading of six related minichromosome maintenance proteins (Mcm2-7) into prereplicative complexes (pre-RCs). Mcm2-7 forms the core of the replicative DNA helicase, which is inactive in the pre-RC. The ATP-dependent Mcm2-7 loading reaction requires the Origin Recognition Complex (ORC), Cdc6 and Cdt1. We have reconstituted Mcm2-7 loading with purified budding yeast proteins. Using biochemical approaches and electron microscopy, we show that single heptamers of Cdt1·Mcm2-7 are loaded cooperatively into stable, head-to-head Mcm2-7 double hexamers connected via N-terminal rings. DNA runs through a central channel in the double hexamer, and, once loaded, Mcm2-7 can slide passively along double-stranded DNA. Our work has significant implications for understanding how eukaryotic DNA replication origins are chosen and licensed, how replisomes assemble during initiation and how unwinding occurs during DNA replication.
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Citations
Cited by
Mechanisms for initiating cellular DNA replication
Bleichert, F., Botchan, M. R., & Berger, J. M. (2017) · Science · doi:10.1126/science.aah6317 · Cited by 220
Replication of G Quadruplex DNA
Lerner, L. K., & Sale, J. E. (2019) · Genes · doi:10.3390/genes10020095 · Cited by 156
LINE-1 protein localization and functional dynamics during the cell cycle
Mita, P., Wudzinska, A., Sun, X., Andrade, J., Nayak, S., Kahler, D. J., Badri, S., LaCava, J., Ueberheide, B., Yun, C. Y., et al. (2018) · eLife · doi:10.7554/elife.30058 · Cited by 142
USP7 Is a Master Regulator of Genome Stability
Valles, G. J., Bezsonova, I., Woodgate, R., & Ashton, N. W. (2020) · Frontiers in Cell and Developmental Biology · doi:10.3389/fcell.2020.00717 · Cited by 114
“Replication licensing is mediated by the licensing factors Cdc6 and Cdt1, which help to load the MCM proteins onto DNA (Evrin et al., 2009; Remus et al., 2009).”
BackgroundNuclear PGK1 Alleviates ADP‐Dependent Inhibition of CDC7 to Promote DNA Replication
Li, X., Qian, X., Jiang, H., Xia, Y., Zheng, Y., Li, J., Huang, B.-J., Fang, J., Qian, C.-N., Jiang, T., et al. (2018) · Molecular Cell · doi:10.1016/j.molcel.2018.09.007 · Cited by 105
Metadata
- Authors
- Remus, D., Beuron, F., Tolun, G., Griffith, J. D., Morris, E. P., & Diffley, J. F. X.
- Venue
- Cell
- Publisher
- Elsevier BV
- Published
- 2009-11
- Volume
- 139
- Issue
- 4
- Pages
- 719-730
- ISSN
- 0092-8674 (ISSN-L)
- OpenAlex
- W2099487576
- Cited by
- 672
- References
- 0
- FWCI
- 14.76
- Type
- journal article
- Language
- en
- Open access
- bronze
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- Remus, D., Beuron, F., Tolun, G., Griffith, J. D., Morris, E. P., & Diffley, J. F. X. (2009). Concerted Loading of Mcm2–7 Double Hexamers around DNA during DNA Replication Origin Licensing. Cell. https://doi.org/10.1016/j.cell.2009.10.015
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