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The S-M checkpoint is an intracellular signaling pathway that ensures that mitosis is not initiated in cells undergoing DNA replication. We identified cid1, a novel fission yeast gene, through its ability when overexpressed to confer specific resistance to a combination of hydroxyurea, which inhibits DNA replication, and caffeine, which overrides the S-M checkpoint. Cid1 overexpression also partially suppressed the hydroxyurea sensitivity characteristic of DNA polymerase delta mutants and mutants defective in the "checkpoint Rad" pathway. Cid1 is a member of a family of putative nucleotidyltransferases including budding yeast Trf4 and Trf5, and mutation of amino acid residues predicted to be essential for this activity resulted in loss of Cid1 function in vivo. Two additional Cid1-like proteins play similar but nonredundant checkpoint-signaling roles in fission yeast. Cells lacking Cid1 were found to be viable but specifically sensitive to the combination of hydroxyurea and caffeine and to be S-M checkpoint defective in the absence of Cds1. Genetic data suggest that Cid1 acts in association with Crb2/Rhp9 and through the checkpoint-signaling kinase Chk1 to inhibit unscheduled mitosis specifically when DNA polymerase delta or epsilon is inhibited.

Type

Journal article

Journal

Mol Cell Biol

Publication Date

05/2000

Volume

20

Pages

3234 - 3244

Keywords

Amino Acid Sequence, Base Sequence, Caffeine, Cell Cycle Proteins, DNA Polymerase II, DNA Polymerase III, DNA-Directed DNA Polymerase, Databases, Factual, Fungal Proteins, Gene Deletion, Genes, cdc, Hydroxyurea, Mitosis, Molecular Sequence Data, Mutagenesis, Site-Directed, Mutation, Nucleic Acid Synthesis Inhibitors, Nucleotidyltransferases, Phosphodiesterase Inhibitors, Phosphoprotein Phosphatases, Plasmids, S Phase, Schizosaccharomyces, Schizosaccharomyces pombe Proteins, Sequence Homology, Amino Acid, Time Factors