Complementation of fission yeast cdc2ts and cdc25ts mutants identifies two cell cycle genes from Drosophila: a cdc2 homologue and string.
We have exploited the universality of the molecular mechanisms that control entry into mitosis to clone the Drosophila melanogaster homologues of fission yeast Schizosaccharomyces pombe cell division control (cdc) genes by the complementation of temperature sensitive mutations. The Drosophila genes were expressed in S.pombe as cDNAs from the SP6 promoter. Successful recovery of complementing plasmids required that we first 'adapt' pooled plasmids from a Drosophila embryonic cDNA library for propagation in fission yeast by introducing an ars1-LEU2 DNA fragment into the vector. This library was introduced into S.pombe cdc2 and cdc25 mutants, and plasmids isolated carrying cDNAs that complement these mutations. The gene that encodes the Drosophila cdc2 homologue maps to a single locus in the Drosophila genome at 31E on chromosome 2. It is expressed maternally to provide mRNA in syncytial embryos, and appears to be zygotically expressed in mitotically active regions of the cellularized embryo. We have isolated two different cDNAs that complement cdc25-22. One corresponds to a transcript of string, previously described as the Drosophila homologue of cdc25, and the other to a gene that has not been previously characterized.
Jimenez
J
J
Department of Biochemistry, University of Oxford, UK.
Alphey
L
L
Nurse
P
P
Glover
D M
DM
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016203:CDC2 Protein Kinase / Q000235:genetics*; D002453:Cell Cycle; D018797:Cell Cycle Proteins; D004247:DNA / Q000235:genetics; D004331:Drosophila melanogaster / Q000196:embryology / Q000235:genetics*; D005656:Fungal Proteins / Q000235:genetics*; D015870:Gene Expression; D005796:Genes; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D012568:Schizosaccharomyces / Q000235:genetics; D020666:ras-GRF1
D018797:Cell Cycle Proteins; D005656:Fungal Proteins; D020666:ras-GRF1; D004247:DNA; D016203:CDC2 Protein Kinase