Dominant mutants identify new roles for p34cdc2 in mitosis.
A large number of dominant mutants have been generated in the fission yeast cdc2 gene, causing lethality when expressed in wild-type cells. The mutants interfere with distinct aspects of p34cdc2 function, producing one of four different phenotypes: mitotic arrest, multiple rounds of S phase in the absence of mitosis, premature mitosis or G2 arrest. The mitotic mutants DL41, DL45 and DL50 are characterized in this paper. Over-expression of DL41 or DL45 causes mitotic arrest, specifically interfering with sister chromatid separation, without preventing spindle elongation. This suggests a role for p34cdc2 in triggering sister chromatid separation at anaphase. DL41 and DL45 also cause abnormal septum formation, suggesting that p34cdc2 may also be involved in regulating this process in fission yeast. These mitotic aspects of p34cdc2 function may involve interaction with p13suc1, since increased expression of suc1 partially suppresses DL41 and DL45. Over-expression of DL50 causes premature mitotic entry in cells that have not completed S phase, resulting in lethality. DL41, DL45 and DL50 correspond to mutation of p34cdc2 residues predicted to be on the surface of the protein, identifying potential sites of interaction with mitotic regulators of p3cdc2, and these residues are conserved amongst cdc2 proteins found in other eukaryotes.
Labib
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Department of Biochemistry, University of Oxford, London, UK.
Craven
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Crawford
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Nurse
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D016203:CDC2 Protein Kinase / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D002842:Chromatids; D015825:Chromosomes, Fungal / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics; D005800:Genes, Fungal / Q000235:genetics*; D005804:Genes, Lethal / Q000235:genetics; D005809:Genes, Regulator / Q000235:genetics; D008938:Mitosis / Q000235:genetics* / Q000502:physiology; D009154:Mutation; D010641:Phenotype; D011401:Promoter Regions, Genetic / Q000235:genetics; D012568:Schizosaccharomyces / Q000235:genetics* / Q000502:physiology; D029702:Schizosaccharomyces pombe Proteins; D013489:Suppression, Genetic
D005656:Fungal Proteins; C093953:NMT1 protein, S pombe; D029702:Schizosaccharomyces pombe Proteins; D016203:CDC2 Protein Kinase