The dynamics of eukaryotic replication initiation: Origin specificity, licensing, and firing at the single-molecule level
Open accessAbstract
Eukaryotic replication initiation is highly regulated and dynamic. It begins with the origin recognition complex (ORC) binding DNA sites called origins of replication. ORC, together with Cdc6 and Cdt1, mediate pre-replicative complex (pre-RC) assembly by loading a double hexamer of Mcm2-7: the core of the replicative helicase. Here, we use single-molecule imaging to directly visualize Saccharomyces cerevisiae pre-RC assembly and replisome firing in real time. We show that ORC can locate and stably bind origins within large tracts of non-origin DNA and that Cdc6 drives ordered pre-RC assembly. We further show that the dynamics of the ORC-Cdc6 interaction dictate Mcm2-7 loading specificity and that Mcm2-7 double hexamers form preferentially at a native origin sequence. Finally, we demonstrate that single Mcm2-7 hexamers propagate bidirectionally, monotonically, and processively as constituents of active replisomes.
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After initial loss of proliferation capacity, a fraction of p21-overexpressing cells evaded arrest/senescence, displaying upregulation of CDT1, CDC6, and ORC.
“After the initial loss of proliferation capacity, a fraction of p21-overexpressing cells evaded arrest/senescence, displaying upregulation of CDT1, CDC6, and ORC, which are all regulators of replication origin licensing [74].”
From CDK-Independent and PCNA-Dependent Functions of p21 in DNA Replication (2020) · cites this paper as [74]
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- Authors
- Duzdevich, D., Warner, M. D., Ticau, S., Ivica, N. A., Bell, S. P., & Greene, E. C.
- Affiliations
- Columbia University (US)
- Massachusetts Institute of Technology (US)
- Howard Hughes Medical Institute (US)
- Corresponding author
- Greene, Eric C.
- Venue
- Molecular Cell
- Publisher
- Elsevier BV
- Published
- 2015-05
- Volume
- 58
- Issue
- 3
- Pages
- 483-494
- ISSN
- 1097-2765 (ISSN-L)
- PMC
- PMC4427541
- OpenAlex
- W2031997435
- Cited by
- 81
- References
- 43
- Citations by year
- 2025: 1 · 2024: 5 · 2023: 7 · 2022: 6 · 2021: 7 · 2020: 7 · 2019: 9 · 2018: 8 · 2017: 15 · 2016: 11 · 2015: 5
- FWCI
- 5.65
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Molecular Cell (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- DSpace@MIT (Massachusetts Institute of Technology) (submittedVersion, cc-by-nc-nd)
- PMC
- Funders
- Howard Hughes Medical Institute
- National Cancer Institute (P30CA014051)
- National Institute of General Medical Sciences (R01GM082848, GM52339, R01GM052339, T32GM007287, GM082848, GM007287)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Microtubule and mitosis dynamics
- Keywords
- origin recognition complex, origin of replication, CDC6, Cdt1, Saccharomyces cerevisiae, single-molecule imaging
- MeSH terms
- Algorithms, Binding Sites, DNA Replication, DNA-Binding Proteins, DNA, Fungal, Eukaryotic Cells, Kinetics, Models, Genetic, Protein Binding, Saccharomyces cerevisiae, Transcription Factors, Replication Origin, Cell Cycle Proteins, Saccharomyces cerevisiae Proteins, Origin Recognition Complex, Minichromosome Maintenance Proteins
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