Journal article
Regulation by phosphorylation of Pho81p, a cyclin-dependent kinase inhibitor in Saccharomyces cerevisiae
Current genetics, v 46(1), pp 10-19
Jul 2004
PMID: 15127225
Abstract
Cyclins, cyclin-dependent kinases (CDK) and their inhibitors play a critical role in many biological processes. In yeast, the ankyrin repeat protein Pho81p, by being an inhibitor of the Pho85p–Pho80p cyclin-dependent protein kinase complex, transcriptionally regulates the production of repressible acid phosphatase, encoded by the PHO5 gene. Recent studies in our laboratory showed that Pho81p is phosphorylated by the Pho80p–Pho85p CDK complex in vitro; and, to determine the significance of the phosphorylation, we used site-directed mutagenesis to alter the potential phosphorylation sites for this kinase complex. The resulting mutations were introduced into a yeast strain containing a deletion of PHO81 and the effect of the mutation on PHO5 expression was assayed. Results suggest that phosphorylation of particular residues within Pho81p is crucial for its activity as an inhibitor. Studies using a green fluorescent protein–Pho81p fusion and Western analysis indicate that the localization and half-life of the mutants are similar to wild-type Pho81 proteins. However, an in vivo binding assay indicates that the mutant Pho81p is deficient in binding to the Pho80p–Pho85p kinase complex. These findings support the observation that the mutant fails to inhibit kinase activity in low phosphate. These studies provide insight into the mechanism of regulation of CDK inhibitor activity.
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Details
- Title
- Regulation by phosphorylation of Pho81p, a cyclin-dependent kinase inhibitor in Saccharomyces cerevisiae
- Creators
- Janine Knight - Mouse Cancer Genetics Program NCI-Frederick Frederick MD 21702-1201 USAThomas Daly - Department of Microbiology and Immunology Drexel University College of Medicine 2900 Queen Lane Philadelphia PA 19129 USALawrence Bergman - Department of Microbiology and Immunology Drexel University College of Medicine 2900 Queen Lane Philadelphia PA 19129 USA
- Publication Details
- Current genetics, v 46(1), pp 10-19
- Publisher
- Springer-Verlag; Berlin/Heidelberg
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Microbiology and Immunology; [Retired Faculty]
- Web of Science ID
- WOS:000222240300002
- Scopus ID
- 2-s2.0-4043181154
- Other Identifier
- 991014877765304721
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- Web of Science research areas
- Genetics & Heredity