Characterization of Schizosaccharomyces pombe mcm7(+) and cdc23(+) (MCM10) and interactions with replication checkpoints.
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 Deltacds1, Deltachk1, or Deltarad3, 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.
Mesh Terms:
Alleles, Base Sequence, Cell Cycle Proteins, Cloning, Molecular, DNA Primers, DNA Replication, Fluorescent Antibody Technique, Indirect, Peptide Initiation Factors, Phenotype, S Phase, Saccharomyces cerevisiae Proteins, Schizosaccharomyces, Temperature, Ubiquitin-Protein Ligase Complexes
Alleles, Base Sequence, Cell Cycle Proteins, Cloning, Molecular, DNA Primers, DNA Replication, Fluorescent Antibody Technique, Indirect, Peptide Initiation Factors, Phenotype, S Phase, Saccharomyces cerevisiae Proteins, Schizosaccharomyces, Temperature, Ubiquitin-Protein Ligase Complexes
Genetics
Date: Oct. 01, 2001
PubMed ID: 11606526
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