Back to Search
Start Over
Stability, chromatin association and functional activity of mammalian pre-replication complex proteins during the cell cycle
- Source :
- The EMBO Journal. 20:4263-4277
- Publication Year :
- 2001
- Publisher :
- Wiley, 2001.
-
Abstract
- We have examined the behavior of pre-replication complex (pre-RC) proteins in relation to key cell cycle transitions in Chinese Hamster Ovary (CHO) cells. ORC1, ORC4 and Cdc6 were stable (T1/22 h) and associated with a chromatin-containing fraction throughout the cell cycle. Green fluorescent protein-tagged ORC1 associated with chromatin throughout mitosis in living cells and co-localized with ORC4 in metaphase spreads. Association of Mcm proteins with chromatin took place during telophase, approximately 30 min after the destruction of geminin and cyclins A and B, and was coincident with the licensing of chromatin to replicate in geminin-supplemented Xenopus egg extracts. Neither Mcm recruitment nor licensing required protein synthesis throughout mitosis. Moreover, licensing could be uncoupled from origin specification in geminin-supplemented extracts; site-specific initiation within the dihydrofolate reductase locus required nuclei from cells that had passed through the origin decision point (ODP). These results demonstrate that mammalian pre-RC assembly takes place during telophase, mediated by post-translational modifications of pre-existing proteins, and is not sufficient to select specific origin sites. A subsequent, as yet undefined, step selects which pre-RCs will function as replication origins.
- Subjects :
- DNA Replication
Origin Recognition Complex
Mitosis
Cell Cycle Proteins
CHO Cells
Xenopus Proteins
Biology
Pre-replication complex
Article
General Biochemistry, Genetics and Molecular Biology
S Phase
Xenopus laevis
Cricetinae
Animals
Humans
Telophase
ORC1
Cell Cycle Protein
Molecular Biology
Mammals
General Immunology and Microbiology
General Neuroscience
Cell Cycle
G1 Phase
Geminin
Minichromosome Maintenance Complex Component 3
Nuclear Proteins
Chromatin
Cell biology
DNA-Binding Proteins
biology.protein
Origin recognition complex
Anaphase
Subjects
Details
- ISSN :
- 14602075
- Volume :
- 20
- Database :
- OpenAIRE
- Journal :
- The EMBO Journal
- Accession number :
- edsair.doi.dedup.....4de5a47ebcb5c3f2590288dc704fb0f6