Characterization of Schizosaccharomyces pombe mcm7(+) and cdc23(+) (MCM10) and interactions with replication checkpoints.

Department of Biology, University of California, San Diego, 92093, USA.
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
Genetics Oct. 01, 2001; 159(2);471-86 [PUBMED:11606526]
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