Characterization of Schizosaccharomyces pombe mcm7⁺ and cdc23⁺ (MCM10) and Interactions With Replication Checkpoints
- Creators
- Liang, Debbie T.
-
Forsburg, Susan L.
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.
Additional Information
© 2001 by the Genetics Society of America. Manuscript received April 25, 2001; Accepted for publication July 12, 2001. We thank Tony Carr, Tamar Enoch, Kathy Gould, Bea Grallert, Dominic Griffiths, Paul Russell, and Rob West for strains, Sally Pasion for isolating the cdc23⁺ genomic clone, Jeff Hodson for tagging and integrating the cdc23HA construct, Ray Chuang for chromatin digestion conditions, Nick Boddy for GST-Weel, and Jill Meissenholder and Tony Hunter for the 12CA5 antibody. We are grateful to Julie Bailis, Mike Catlett, Eliana Gomez, Sally Pasion, and Nick Rhind for critical reading of the manuscript, and all Forsburg laboratory members for helpful discussions. This work was supported by National Institutes of Health grant GM-59321 to S.L.F., who is a scholar of the Leukemia and Lymphoma Society.Attached Files
Published - 471.full.pdf
Files
| Name | Size | Download all |
|---|---|---|
|
md5:1616eb56191a099d61a1c2768821569f
|
450.6 kB | Preview Download |
Additional details
- Alternative title
- Characterization of Schizosaccharomyces pombe mcm7+ and cdc23+ (MCM10) and Interactions With Replication Checkpoints
- PMCID
- PMC1461838
- Eprint ID
- 108492
- Resolver ID
- CaltechAUTHORS:20210318-150836527
- GM-59321
- NIH
- Leukemia and Lymphoma Society
- Created
-
2021-03-19Created from EPrint's datestamp field
- Updated
-
2021-03-19Created from EPrint's last_modified field