Characterization of Schizosaccharomyces pombe mcm7⁺ and cdc23⁺ (MCM10) and Interactions With Replication Checkpoints

Corresponding author: Susan L. Forsburg, MCBL-F, The Salk Institute, 10010 N. Torrey Pines Rd., La Jolla, CA 92037., forsburg{at}salk.edu (E-mail)

Communicating editor: P. RUSSELL

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 cds1, chk1, or rad3, 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.

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