A fission yeast strain expressing human CDC25A phosphatase: a tool for selectivity studies of pharmacological inhibitors of CDC25.
Fission yeast is a simple eukaryotic model organism in which many aspects of cell cycle control can be explored. We examined by homologous recombination whether the human CDC25A phosphatase could substitute for the function of the fission yeast Cdc25. We first show: (a). that CDC25A efficiently replaces the endogenous Cdc25 mitotic inducer for vegetative growth and (b). that CDC25A is able to partially restore a functional checkpoint in response to both ionising and UV irradiation, but not a DNA replication checkpoint. We then describe a simple assay in which we demonstrate that growth of the humanised CDC25A strain is strongly repressed in a CDC25-dependent manner by BN2003, a potent chemical inhibitor of CDC25 belonging to the benzothiazoledione family. The ease of manipulation of fission yeast humanised CDC25 cells and the simplicity of the above assay offer a powerful tool with which to investigate the specificity of pharmacological inhibitors of CDC25.
Mondesert
Odile
O
LBCMCP-CNRS UMR5088, Université Paul Sabatier, IFR109 "Institut d'Exploration Fonctionnelle des Génomes", 118 route de Narbonne, 31077, Toulouse, France.
Lemaire
Matthieu
M
Brezak
Marie-Christine
MC
Galera-Contour
Marie Odile
MO
Prevost
Grégoire
G
Ducommun
Bernard
B
Bugler
Béatrix
B
D052160:Benzothiazoles; D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors*; D049109:Cell Proliferation; D004247:DNA / Q000378:metabolism; D004249:DNA Damage; D004305:Dose-Response Relationship, Drug; D004307:Dose-Response Relationship, Radiation; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005656:Fungal Proteins / Q000037:antagonists & inhibitors*; D005838:Genotype; D006801:Humans; D006918:Hydroxyurea / Q000494:pharmacology; D007259:Infrared Rays; D020128:Inhibitory Concentration 50; D008938:Mitosis; D008957:Models, Genetic; D010744:Phosphoric Monoester Hydrolases / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D020033:Protein Isoforms; D012568:Schizosaccharomyces / Q000378:metabolism*; D013696:Temperature; D013844:Thiazoles / Q000737:chemistry; D013997:Time Factors; D014466:Ultraviolet Rays; D020687:cdc25 Phosphatases / Q000737:chemistry* / Q000378:metabolism*; D020666:ras-GRF1 / Q000037:antagonists & inhibitors*
D052160:Benzothiazoles; D018797:Cell Cycle Proteins; D004791:Enzyme Inhibitors; D005656:Fungal Proteins; D020033:Protein Isoforms; D013844:Thiazoles; D020666:ras-GRF1; C058836:benzothiazoline; D004247:DNA; D010744:Phosphoric Monoester Hydrolases; C517004:CDC25A protein, human; D020687:cdc25 Phosphatases; D006918:Hydroxyurea