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Localization and regulation of the cdk-activating kinase (Cak1p) from budding yeast

Kaldis, P. LU orcid ; Pitluk, Z. W. ; Bany, I. A. ; Enke, D. A. ; Wagner, M. ; Winter, E. and Solomon, M. J. (1998) In Journal of Cell Science 111(24). p.3585-3596
Abstract

Eukaryotic cell cycles are controlled by the activities of cyclin-dependent kinases (cdks). The major cdk in budding yeast, Saccharomyces cerevisiae, is Cdc28p. Activation of Cdc28p requires phosphorylation on threonine 169 and binding to a cyclin. Thr-169 is phosphorylated by the cdk-activating kinase (CAK), Cak1p, which was recently identified as the physiological CAK in budding yeast. Here we present our further characterization of yeast Cak1p. We have found that Cak1p is dispersed throughout the cell as shown by immunofluorescence; biochemical subcellular fractionation confirmed that most of the Cak1p is found in the cytoplasm. Cak1p is a monomeric enzyme in crude yeast lysates. Mutagenesis of potential sites of activating... (More)

Eukaryotic cell cycles are controlled by the activities of cyclin-dependent kinases (cdks). The major cdk in budding yeast, Saccharomyces cerevisiae, is Cdc28p. Activation of Cdc28p requires phosphorylation on threonine 169 and binding to a cyclin. Thr-169 is phosphorylated by the cdk-activating kinase (CAK), Cak1p, which was recently identified as the physiological CAK in budding yeast. Here we present our further characterization of yeast Cak1p. We have found that Cak1p is dispersed throughout the cell as shown by immunofluorescence; biochemical subcellular fractionation confirmed that most of the Cak1p is found in the cytoplasm. Cak1p is a monomeric enzyme in crude yeast lysates. Mutagenesis of potential sites of activating phosphorylation had little effect on the activity of Cak1p in vitro or in vivo. Furthermore, Cak1p contains no posttranslational modifications detectable by two-dimensional isoelectric focusing. We found that Cak1p is a stable protein during exponential growth but that its expression decreases considerably when cells enter stationary phase. In contrast, Cak1p levels oscillate dramatically during meiosis, reflecting regulation at both the transcriptional and post-translational level. The localization and regulation of Cak1p are in contrast to those of the known vertebrate CAK, p40(MO15).

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author
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publishing date
type
Contribution to journal
publication status
published
keywords
CAK, Cak1p, cdk, Cyclin-dependent kinase, Meiosis, Yeast
in
Journal of Cell Science
volume
111
issue
24
pages
3585 - 3596
publisher
The Company of Biologists Ltd
external identifiers
  • scopus:0032433028
  • pmid:9819350
ISSN
0021-9533
language
English
LU publication?
no
id
6239b380-a6d7-424f-a6a2-962e411e02a6
date added to LUP
2019-09-18 14:35:10
date last changed
2024-06-27 06:05:51
@article{6239b380-a6d7-424f-a6a2-962e411e02a6,
  abstract     = {{<p>Eukaryotic cell cycles are controlled by the activities of cyclin-dependent kinases (cdks). The major cdk in budding yeast, Saccharomyces cerevisiae, is Cdc28p. Activation of Cdc28p requires phosphorylation on threonine 169 and binding to a cyclin. Thr-169 is phosphorylated by the cdk-activating kinase (CAK), Cak1p, which was recently identified as the physiological CAK in budding yeast. Here we present our further characterization of yeast Cak1p. We have found that Cak1p is dispersed throughout the cell as shown by immunofluorescence; biochemical subcellular fractionation confirmed that most of the Cak1p is found in the cytoplasm. Cak1p is a monomeric enzyme in crude yeast lysates. Mutagenesis of potential sites of activating phosphorylation had little effect on the activity of Cak1p in vitro or in vivo. Furthermore, Cak1p contains no posttranslational modifications detectable by two-dimensional isoelectric focusing. We found that Cak1p is a stable protein during exponential growth but that its expression decreases considerably when cells enter stationary phase. In contrast, Cak1p levels oscillate dramatically during meiosis, reflecting regulation at both the transcriptional and post-translational level. The localization and regulation of Cak1p are in contrast to those of the known vertebrate CAK, p40(MO15).</p>}},
  author       = {{Kaldis, P. and Pitluk, Z. W. and Bany, I. A. and Enke, D. A. and Wagner, M. and Winter, E. and Solomon, M. J.}},
  issn         = {{0021-9533}},
  keywords     = {{CAK; Cak1p; cdk; Cyclin-dependent kinase; Meiosis; Yeast}},
  language     = {{eng}},
  month        = {{12}},
  number       = {{24}},
  pages        = {{3585--3596}},
  publisher    = {{The Company of Biologists Ltd}},
  series       = {{Journal of Cell Science}},
  title        = {{Localization and regulation of the cdk-activating kinase (Cak1p) from budding yeast}},
  volume       = {{111}},
  year         = {{1998}},
}