title: Yeast as a model system to screen purine derivatives against human CDK1 and CDK2 kinases.
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.
authors.affiliation: INSERM U612, Institut Curie, Bât. 110-112, Centre Universitaire, 91405 Orsay, France.
authors.forename: Thérèse
authors.initials: T
authors.lastname: Mayi
authors.affiliation: CNRS UMR2027, Institut Curie, Bât. 110-112, Centre Universitaire, 91405 Orsay, France.
authors.forename: Céline
authors.initials: C
authors.lastname: Facca
authors.affiliation: CNRS UMR146, Institut Curie, Bât. 110-112, Centre Universitaire, 91405 Orsay, France.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Anne
authors.affiliation: CNRS UMR3348, Institut Curie, Bât. 110-112, Centre Universitaire, 91405 Orsay, France.
authors.forename: Laurence
authors.initials: L
authors.lastname: Vernis
authors.affiliation: CNRS UMR3348, Institut Curie, Bât. 110-112, Centre Universitaire, 91405 Orsay, France.
authors.forename: Meng-Er
authors.initials: ME
authors.lastname: Huang
authors.affiliation: CNRS UMR176, Institut Curie, Bât. 110-112, Centre Universitaire, 91405 Orsay, France.
authors.forename: Michel
authors.initials: M
authors.lastname: Legraverend
authors.affiliation: CNRS UMR3348, Institut Curie, Bât. 110-112, Centre Universitaire, 91405 Orsay, France. Electronic address: gerard.faye405@orange.fr.
authors.forename: Gérard
authors.initials: G
authors.lastname: Faye
mesh_terms: D016203:CDC2 Protein Kinase / Q000037:antagonists & inhibitors* / Q000235:genetics; D017489:CDC28 Protein Kinase, S cerevisiae / Q000235:genetics; D051357:Cyclin-Dependent Kinase 2 / Q000037:antagonists & inhibitors* / Q000235:genetics; D055808:Drug Discovery / Q000379:methods*; D005818:Genetic Engineering; D006801:Humans; D008954:Models, Biological; D011687:Purines / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology*; D012441:Saccharomyces cerevisiae
chemical_list: D011687:Purines; D016203:CDC2 Protein Kinase; D017489:CDC28 Protein Kinase, S cerevisiae; C495894:CDK2 protein, human; D051357:Cyclin-Dependent Kinase 2
keywords: Cyclin-dependent kinases; In vivo screen; Purine derivatives; Yeast cells