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Sonneville, Remi, Querenet, Matthieu, Craig, Ashley, Gartner, Anton and Blow, J. Julian 
ORCID: https://orcid.org/0000-0002-9524-5849
(2012)
The dynamics of replication licensing in live Caenorhabditis elegans embryos.
Journal of Cell Biology, 196 (2).
pp. 233-246.
ISSN 0021-9525
Abstract
Accurate DNA replication requires proper regulation of replication licensing, which entails loading MCM-2-7 onto replication origins. In this paper, we provide the first comprehensive view of replication licensing in vivo, using video microscopy of Caenorhabditis elegans embryos. As expected, MCM-2-7 loading in late M phase depended on the prereplicative complex (pre-RC) proteins: origin recognition complex (ORC), CDC-6, and CDT-1. However, many features we observed have not been described before: GFP-ORC-1 bound chromatin independently of ORC-2-5, and CDC-6 bound chromatin independently of ORC, whereas CDT-1 and MCM-2-7 DNA binding was interdependent. MCM-3 chromatin loading was irreversible, but CDC-6 and ORC turned over rapidly, consistent with ORC/CDC-6 loading multiple MCM-2-7 complexes. MCM-2-7 chromatin loading further reduced ORC and CDC-6 DNA binding. This dynamic behavior creates a feedback loop allowing ORC/CDC-6 to repeatedly load MCM-2-7 and distribute licensed origins along chromosomal DNA. During S phase, ORC and CDC-6 were excluded from nuclei, and DNA was overreplicated in export-defective cells. Thus, nucleocytoplasmic compartmentalization of licensing factors ensures that DNA replication occurs only once.
| Item Type: | Article |
|---|---|
| Faculty \ School: | |
| Depositing User: | LivePure Connector |
| Date Deposited: | 10 Jun 2024 15:31 |
| Last Modified: | 25 Sep 2024 17:51 |
| URI: | https://ueaeprints.uea.ac.uk/id/eprint/95473 |
| DOI: | 10.1083/jcb.201110080 |
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