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Detection of Homologous Recombination Intermediates via Proximity Ligation and Quantitative PCR in Saccharomyces cerevisiae
Published on: September 11, 2022
Sld3, which interacts with Cdc45 (Sld4), functions for chromosomal DNA replication in Saccharomyces cerevisiae
Y Kamimura1, Y S Tak, A Sugino
1Division of Microbial Genetics, National Institute of Genetics, Osaka University, Osaka 565-0871 Japan.
The EMBO Journal
|April 11, 2001
Summary
The novel protein Sld3 forms a complex with Cdc45, crucial for DNA replication initiation. This Sld3-Cdc45 complex is essential for recruiting replication machinery to origins, enabling DNA unwinding.
Area of Science:
- Molecular Biology
- Cell Biology
- Genetics
Background:
- Cdc45 is vital for eukaryotic DNA replication initiation and elongation.
- Cdc45 interacts with minichromosomal maintenance (Mcm) proteins.
Purpose of the Study:
- To investigate the role of a novel factor, Sld3, in complex formation with Cdc45.
- To elucidate the function of the Sld3-Cdc45 complex in DNA replication.
Main Methods:
- Chromatin immunoprecipitation assays in Saccharomyces cerevisiae.
- Analysis of temperature-sensitive sld3 mutations.
- Assessment of replication factor A association with origins.
Main Results:
- Sld3 forms a stable complex with Cdc45 throughout the cell cycle.
- Both Sld3 and Cdc45 bind simultaneously and dependently to replication origins.
- The Sld3-Cdc45 complex interacts with Mcm proteins, and its origin association precedes DNA unwinding.
Conclusions:
- The Sld3-Cdc45 complex is a key component in recruiting replication machinery to origins.
- Origin association of the Sld3-Cdc45 complex is a prerequisite for DNA unwinding during replication initiation.
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