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Fission yeast Cdc23 interactions with DNA replication initiation proteins.
Elizabeth A Hart1, John A Bryant, Karen Moore
1School of Biological Sciences, University of Exeter, Perry Road, Exeter EX4 4QG, UK.
Current Genetics
|August 20, 2002
Summary
Fission yeast Cdc23 interacts with MCM proteins and origin recognition complex, suggesting a role in DNA replication initiation. This essential protein is crucial for DNA replication, interacting with key complexes.
Area of Science:
- Molecular Biology
- Cell Biology
- Genetics
Background:
- Schizosaccharomyces pombe Cdc23 is an essential DNA replication protein.
- It is conserved in eukaryotes and homologous to Saccharomyces cerevisiae Dna43 (Mcm10).
- The MCM2-7 complex is vital for DNA replication initiation and elongation.
Purpose of the Study:
- To investigate the interactions between Cdc23 and the MCM2-7 complex.
- To determine the role of Cdc23 in DNA replication.
- To explore potential interactions with the origin recognition complex (ORC).
Main Methods:
- Genetic interaction analysis using various mutant alleles.
- Yeast two-hybrid assays to detect protein-protein interactions.
- Analysis of Cdc23 mRNA levels throughout the cell cycle.
Main Results:
- Cdc23 exhibits synthetic genetic interactions with multiple MCM subunits (Mcm2, Mcm4, Mcm5, Mcm6).
- Two-hybrid analysis confirmed direct protein-protein interactions between Cdc23 and Mcm4, Mcm5, and Mcm6.
- Cdc23 also interacts with Orc1, Orc2, Orc5, and Orc6 of the ORC, but not with Cdc18.
- Cdc23 mRNA levels remain stable throughout the cell cycle.
Conclusions:
- Fission yeast Cdc23 is an MCM-associated factor.
- Cdc23 plays a role in the initiation of DNA replication.
- Its interactions with MCM and ORC subunits highlight its involvement in replication regulation.