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Updated: Apr 19, 2026
Yeast As a Chassis for Developing Functional Assays to Study Human P53
Published on: August 4, 2019
Yeast as a model system to screen purine derivatives against human CDK1 and CDK2 kinases
Thérèse Mayi1, Céline Facca2, Sandrine Anne3
1INSERM U612, Institut Curie, Bât. 110-112, Centre Universitaire, 91405 Orsay, France.
Abstract:
Cyclin-dependent kinases (Cdk) play crucial roles in cell cycle progression. Aberrant activation of Cdk1 has been observed in a number of primary tumors and Cdk2 is deregulated in various malignancies. The therapeutic value of targeting Cdk1 and Cdk2 has been explored in a number of experimental systems. In the present study, taking advantage of the fact that deletion of the yeast CDC28 gene is functionally complemented by human CDK1 or CDK2, we set up an in vivo screen system to evaluate the inhibitory potency of purine derivatives against these two human Cdks. We constructed three isogenic strains highly sensitive to small molecules and harboring genes CDK1, CDK2 or CDC28, under the control of the CDC28 promoter. In a proof of principle assay, we determined the inhibitory effect of 82 purine derivatives on the growth rate of these strains. Thirty-three of them were revealed to be able to inhibit the Cdk1- or Cdk2-harboring strains but not the Cdc28-harboring strain, suggesting a specific inhibitory effect on human Cdks. Our data demonstrate that the yeast-based assay is an efficient system to identify potential specific inhibitors that should be preferentially selected for further investigation in cultured human cell lines.
Insights
Researchers developed a yeast-based screening system to identify specific inhibitors of human cyclin-dependent kinases (Cdk1 and Cdk2). This method efficiently detects compounds that target cancer-related kinases, aiding drug discovery for malignancies.
Area of Science:
- Molecular Biology
- Biochemistry
- Pharmacology
Background:
- Cyclin-dependent kinases (Cdks) are vital regulators of cell cycle progression.
- Aberrant activity of Cdk1 and Cdk2 is implicated in various cancers, making them therapeutic targets.
- Existing experimental systems for evaluating Cdk inhibitors are limited.
Purpose of the Study:
- To establish and validate a yeast-based in vivo screening system for identifying specific inhibitors of human Cdk1 and Cdk2.
- To evaluate the efficacy of purine derivatives as potential Cdk inhibitors using this novel system.
Main Methods:
- Constructed three isogenic yeast strains sensitive to small molecules, expressing human CDK1, CDK2, or the yeast CDC28 gene under the CDC28 promoter.
- Screened 82 purine derivatives for their ability to inhibit the growth rate of these engineered yeast strains.
- Identified compounds that selectively inhibited human CDK1 or CDK2 but not the yeast Cdc28.
Main Results:
- Thirty-three out of 82 tested purine derivatives demonstrated inhibitory activity against human Cdk1 or Cdk2.
- These 33 compounds did not inhibit the growth of the yeast strain expressing Cdc28, indicating specificity for human Cdks.
- The yeast-based assay proved efficient in identifying potential specific inhibitors.
Conclusions:
- The developed yeast screening system is an effective tool for discovering specific inhibitors of human Cdk1 and Cdk2.
- This method facilitates the selection of promising drug candidates for further investigation in human cell lines.
- The findings support the therapeutic potential of targeting Cdk1 and Cdk2 in cancer treatment.
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