Characterization of Schizosaccharomyces pombe mcm7+ and cdc23+ (MCM10) and Interactions With Replication Checkpoints
Copyright policy )Characterization of Schizosaccharomyces pombe mcm7+ and cdc23+ (MCM10) and Interactions With Replication Checkpoints
Abstract MCM proteins are required for the proper regulation of DNA replication. We cloned fission yeast mcm7+ and showed it is essential for viability; spores lacking mcm7+ begin S phase later than wild-type cells and arrest with an apparent 2C DNA content. We isolated a novel temperature-sensitive allele, mcm7-98, and also characterized two temperature-sensitive alleles of the fission yeast homolog of MCM10, cdc23+. mcm7-98 and both cdc23ts alleles arrest with damaged chromosomes and an S phase delay. We find that mcm7-98 is synthetically lethal with the other mcmts mutants but does not interact genetically with either cdc23ts allele. However, cdc23-M36 interacts with mcm4ts. Unlike other mcm mutants or cdc23, mcm7-98 is synthetically lethal with checkpoint mutants Δcds1, Δchk1, or Δrad3, suggesting chromosomal defects even at permissive temperature. Mcm7p is a nuclear protein throughout the cell cycle, and its localization is dependent on the other MCM proteins. Our data suggest that the Mcm3p-Mcm5p dimer interacts with the Mcm4p-Mcm6p-Mcm7p core complex through Mcm7p.
- University of California, San Diego United States
- California Institute of Technology United States
- University of California, San Diego United States
- Salk Institute for Biological Studies United States
DNA Replication, 570, Saccharomyces cerevisiae Proteins, Base Sequence, Temperature, 610, Ubiquitin-Protein Ligase Complexes, Cell Cycle Proteins, Apc8 Subunit, Anaphase-Promoting Complex-Cyclosome, Anaphase-Promoting Complex-Cyclosome, S Phase, Phenotype, Peptide Initiation Factors, Schizosaccharomyces, Cloning, Molecular, Fluorescent Antibody Technique, Indirect, Alleles, DNA Primers
DNA Replication, 570, Saccharomyces cerevisiae Proteins, Base Sequence, Temperature, 610, Ubiquitin-Protein Ligase Complexes, Cell Cycle Proteins, Apc8 Subunit, Anaphase-Promoting Complex-Cyclosome, Anaphase-Promoting Complex-Cyclosome, S Phase, Phenotype, Peptide Initiation Factors, Schizosaccharomyces, Cloning, Molecular, Fluorescent Antibody Technique, Indirect, Alleles, DNA Primers
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