Loss of a protein phosphatase 2A regulatory subunit (Cdc55p) elicits improper regulation of Swe1p degradation.
CDC55 encodes a Saccharomyces cerevisiae protein phosphatase 2A (PP2A) regulatory subunit. cdc55-null cells growing at low temperature exhibit a failure of cytokinesis and produce abnormally elongated buds, but cdc55-null cells producing the cyclin-dependent kinase Cdc28-Y19F, which is unable to be inhibited by Y19 phosphorylation, show a loss of the abnormal ... morphology. Furthermore, cdc55-null cells exhibit a hyperphosphorylation of Y19. For these reasons, we have examined in wild-type and cdc55-null cells the levels and activities of the kinase (Swe1p) and phosphatase (Mih1p) that normally regulate the extent of Cdc28 Y19 phosphorylation. We find that Mih1p levels are comparable in the two strains, and an estimate of the in vivo and in vitro phosphatase activity of this enzyme in the two cell types indicates no marked differences. By contrast, while Swe1p levels are similar in unsynchronized and S-phase-arrested wild-type and cdc55-null cells, Swe1 kinase is found at elevated levels in mitosis-arrested cdc55-null cells. This excess Swe1p in cdc55-null cells is the result of ectopic stabilization of this protein during G(2) and M, thereby accounting for the accumulation of Swe1p in mitosis-arrested cells. We also present evidence indicating that, in cdc55-null cells, misregulated PP2A phosphatase activity is the cause of both the ectopic stabilization of Swe1p and the production of the morphologically abnormal phenotype.
Mesh Terms:
Alkaline Phosphatase, Antineoplastic Agents, Blotting, Northern, Blotting, Western, Cell Cycle Proteins, Cell Division, Electrophoresis, Polyacrylamide Gel, Fungal Proteins, G2 Phase, Gene Deletion, Kinetics, Mitosis, Nocodazole, Phenotype, Phosphoprotein Phosphatases, Phosphorylation, Plasmids, Protein Phosphatase 2, Protein-Tyrosine Kinases, RNA, Messenger, RNA, Ribosomal, S Phase, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Temperature, Time Factors, cdc25 Phosphatases, ras-GRF1
Alkaline Phosphatase, Antineoplastic Agents, Blotting, Northern, Blotting, Western, Cell Cycle Proteins, Cell Division, Electrophoresis, Polyacrylamide Gel, Fungal Proteins, G2 Phase, Gene Deletion, Kinetics, Mitosis, Nocodazole, Phenotype, Phosphoprotein Phosphatases, Phosphorylation, Plasmids, Protein Phosphatase 2, Protein-Tyrosine Kinases, RNA, Messenger, RNA, Ribosomal, S Phase, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Temperature, Time Factors, cdc25 Phosphatases, ras-GRF1
Mol. Cell. Biol.
Date: Nov. 01, 2000
PubMed ID: 11027284
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