Genetic analysis of human p34CDC2 function in fission yeast.
The p34cdc2 protein kinase plays a key role in the control of the mitotic cell cycle of fission yeast, being required for both entry into S-phase and for entry into mitosis in the mitotic cell cycle, as well as for the initiation of the second meiotic nuclear division. In recent years, structural and functional homologues of p34cdc2, as well as several of the proteins that interact with and regulate p34cdc2 function in fission yeast, have been identified in a wide range of higher eukaryotic cell types, suggesting that the control mechanisms uncovered in this simple eukaryote are likely to be well conserved across evolution. Here we describe the construction and characterisation of a fission yeast strain in which the endogenous p34cdc2 protein is entirely absent and is replaced by its human functional homologue p34CDC2. We have used this strain to analyse aspects of the function of the human p34CDC2 protein genetically. We show that the function of the human p34CDC2 protein in fission yeast cells is dependent upon the action of the protein tyrosine phosphatase p80cdc25, that it responds to altered levels of both the mitotic inhibitor p107wee1 and the p34cdc2-binding protein p13suc1, and is lethal in combination with the mutant B-type cyclin p56cdc13-117. In addition, we demonstrate that the human p34CDC2 protein is proficient for fission yeast meiosis, and examine the behaviour of two mutant p34CDC2 proteins in fission yeast.
MacNeill
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SA
Department of Biochemistry, University of Oxford, UK.
Nurse
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D000483:Alleles; D000595:Amino Acid Sequence; D001483:Base Sequence; D016203:CDC2 Protein Kinase / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D018797:Cell Cycle Proteins; D005656:Fungal Proteins / Q000378:metabolism; D005800:Genes, Fungal; D006801:Humans; D008540:Meiosis; D008969:Molecular Sequence Data; D009154:Mutation; D009687:Nuclear Proteins; D009838:Oligodeoxyribonucleotides; D011494:Protein Kinases / Q000378:metabolism; D011505:Protein-Tyrosine Kinases; D012568:Schizosaccharomyces / Q000166:cytology / Q000201:enzymology* / Q000235:genetics; D029702:Schizosaccharomyces pombe Proteins; D017386:Sequence Homology, Amino Acid; D013696:Temperature
D018797:Cell Cycle Proteins; D005656:Fungal Proteins; D009687:Nuclear Proteins; D009838:Oligodeoxyribonucleotides; D029702:Schizosaccharomyces pombe Proteins; C051655:Suc1 protein, S pombe; D011494:Protein Kinases; C068689:wee1 protein, S pombe; D011505:Protein-Tyrosine Kinases; C499349:WEE1 protein, human; D016203:CDC2 Protein Kinase