Book chapter
Combining the Optimized Yeast Cytosine Deaminase Protein Fragment Complementation Assay and an In Vitro Cdk1 Targeting Assay to Study the Regulation of the gamma-Tubulin Complex
Cell Cycle Oscillators, pp.237-257
Methods in Molecular Biology, Humana Press Inc
01/01/2016
DOI: 10.1007/978-1-4939-2957-3_14
PMID: 26254928
Abstract
Cdk1 is the essential cyclin-dependent kinase in the budding yeast Saccharomyces cerevisiae. Cdk1 orchestrates cell cycle control by phosphorylating target proteins with extraordinary temporal and spatial specificity by complexing with one of the nine cyclin regulatory subunits. The identification of the cyclin required for targeting Cdk1 to a substrate can help to place the regulation of that protein at a specific time point during the cell cycle and reveal information needed to elucidate the biological significance of the regulation. Here, we describe a combination of strategies to identify interaction partners of Cdk1, and associate these complexes to the appropriate cyclins using a cell-based protein-fragment complementation assay. Validation of the specific reliance of the OyCD interaction between Cdk1 and budding yeast gamma-tubulin on the Clb3 cyclin, relative to the mitotic Clb2 cyclin, was performed by an in vitro kinase assay using the gamma-tubulin complex as a substrate.
Details
- Title: Subtitle
- Combining the Optimized Yeast Cytosine Deaminase Protein Fragment Complementation Assay and an In Vitro Cdk1 Targeting Assay to Study the Regulation of the gamma-Tubulin Complex
- Creators
- Po Hien Ear - Harvard UniversityJacqueline Kowarzyk - Université de MontréalMichael J. Booth - Université de MontréalDiala Abd-Rabbo - Université de MontréalKristian Shulist - McGill UniversityConrad Hall - McGill UniversityJackie Vogel - McGill UniversityStephen W. Michnick - Université de Montréal
- Contributors
- A S Coutts (Editor)L Weston (Editor)
- Resource Type
- Book chapter
- Publication Details
- Cell Cycle Oscillators, pp.237-257
- Publisher
- Humana Press Inc; TOTOWA
- Series
- Methods in Molecular Biology
- DOI
- 10.1007/978-1-4939-2957-3_14
- PMID
- 26254928
- eISSN
- 1940-6029
- ISSN
- 1064-3745
- Number of pages
- 21
- Language
- English
- Date published
- 01/01/2016
- Academic Unit
- Surgery
- Record Identifier
- 9984322826402771
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