Site-specific mutagenesis of cdc2+, a cell cycle control gene of the fission yeast Schizosaccharomyces pombe.
The cdc2+ gene of Schizosaccharomyces pombe is homologous to the CDC28 gene of Saccharomyces cerevisiae. Both genes share limited homology with vertebrate protein kinases and have protein kinase activity. cdc2+ has been subjected to mutagenesis in vitro. A null allele of the gene, constructed by insertion of the S. cerevisiae LEU2 gene into a site within the gene, has a phenotype similar to that of many temperature-sensitive alleles of cdc2. Mutations within the predicted ATP-binding site and in a region which may be a site of phosphorylation result in loss of cdc2+ activity. A single substitution of Gly-146 to Asp-146 has been identified in cdc2-1w, a dominant activated allele of the gene. The four introns within the cdc2+ gene have been deleted. The resulting gene not only functions in fission yeast but also rescues cdc28(Ts) strains of S. cerevisiae, a property which is not shared by the genomic cdc2+ gene.
D000483:Alleles; D000818:Animals; D001483:Base Sequence; D002453:Cell Cycle; D005796:Genes; D005800:Genes, Fungal; D009154:Mutation; D011494:Protein Kinases / Q000235:genetics*; D004718:Saccharomycetales / Q000235:genetics*; D012568:Schizosaccharomyces / Q000166:cytology / Q000235:genetics* / Q000254:growth & development; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity
D011494:Protein Kinases