title: Redundant mechanisms prevent mitotic entry following replication arrest in the absence of Cdc25 hyper-phosphorylation in fission yeast.
abstract: Following replication arrest the Cdc25 phosphatase is phosphorylated and inhibited by Cds1. It has previously been reported that expressing Cdc25 where 9 putative amino-terminal Cds1 phosphorylation sites have been substituted to alanine results in bypass of the DNA replication checkpoint. However, these results were acquired by expression of the phosphorylation mutant using a multicopy expression vector in a genetic background where the DNA replication checkpoint is intact. In order to clarify these results we constructed a Cdc25(9A)-GFP native promoter integrant and examined its effect on the replication checkpoint at endogenous expression levels. In this strain the replication checkpoint operates normally, conditional on the presence of the Mik1 kinase. In response to replication arrest the Cdc25(9A)-GFP protein is degraded, suggesting the presence of a backup mechanism to eliminate the phosphatase when it cannot be inhibited through phosphorylation.
authors: Department of Biology, Queen's University, Kingston, Ontario, Canada.
authors: Corey
authors: C
authors: Frazer
authors: Paul G
authors: PG
authors: Young
mesh_terms: D004249:DNA Damage; D004261:DNA Replication / Q000187:drug effects; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006918:Hydroxyurea / Q000494:pharmacology; D008938:Mitosis / Q000187:drug effects; D009154:Mutation / Q000235:genetics; D010766:Phosphorylation / Q000187:drug effects; D011401:Promoter Regions, Genetic / Q000235:genetics; D021381:Protein Transport / Q000187:drug effects; D012568:Schizosaccharomyces / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology* / Q000254:growth & development; D029702:Schizosaccharomyces pombe Proteins / Q000378:metabolism*; D020687:cdc25 Phosphatases / Q000378:metabolism*
chemical_list: D029702:Schizosaccharomyces pombe Proteins; D049452:Green Fluorescent Proteins; D020687:cdc25 Phosphatases; D006918:Hydroxyurea