Okadaic acid, a potent inhibitor of type 1 and type 2A protein phosphatases, activates cdc2/H1 kinase and transiently induces a premature mitosis-like state in BHK21 cells.
When BHK21 cells synchronized in early S phase were exposed to okadaic acid (OA), an inhibitor of protein phosphatases 1 and 2A, mitosis specific events such as premature chromosome condensation, the production of MPM-2 antigens, dispersion of nuclear lamins and the appearance of mitotic asters were induced, and then disappeared upon further incubation. These mitosis specific events occurred even in the presence of cycloheximide. Within 1 h of exposure to OA, cdc2/histone H1 kinase activity rose 10-fold compared with untreated controls, but returned to the control level upon further incubation. Using antibodies against either p34cdc2 or cyclin B it was found that p34cdc2 complexed with cyclin B was dephosphorylated after OA treatment concomitant with the activation of cdc2 kinase, and that cyclin B was subsequently degraded concomitant with a decrease in cdc2 kinase activity, as in normal mitosis. In contrast, when cells in G1 phase were treated with OA no increase in cdc2 kinase activity was observed. Moreover when cells in pseudo-metaphase induced by nocodazole were treated with OA, cdc2 kinase was inactivated. These results suggest that OA sensitive protein phosphatases control both the activation and inactivation of the p34cdc2 kinase.
Yamashita
K
K
Department of Molecular Biology, Graduate School of Medical Science, Kyushu University, Fukuoka, Japan.
Yasuda
H
H
Pines
J
J
Yasumoto
K
K
Nishitani
H
H
Ohtsubo
M
M
Hunter
T
T
Sugimura
T
T
Nishimoto
T
T
D000818:Animals; D016203:CDC2 Protein Kinase / Q000096:biosynthesis* / Q000235:genetics; D002352:Carrier Proteins; D002453:Cell Cycle; D002460:Cell Line; D002875:Chromosomes; D006224:Cricetinae; D016213:Cyclins / Q000378:metabolism; D003513:Cycloheximide / Q000494:pharmacology; D004351:Drug Resistance; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D004988:Ethers, Cyclic / Q000494:pharmacology*; D047908:Intracellular Signaling Peptides and Proteins; D008647:Mesocricetus; D008938:Mitosis; D019319:Okadaic Acid; D010749:Phosphoprotein Phosphatases / Q000378:metabolism*; D011506:Proteins / Q000494:pharmacology*; D014178:Translocation, Genetic; D000097847:Protein Phosphatase Inhibitory Proteins
D016203:CDC2 Protein Kinase; D002352:Carrier Proteins; D016213:Cyclins; D003513:Cycloheximide; D004791:Enzyme Inhibitors; D004988:Ethers, Cyclic; D047908:Intracellular Signaling Peptides and Proteins; D019319:Okadaic Acid; D010749:Phosphoprotein Phosphatases; D011506:Proteins; C021042:protein phosphatase inhibitor-1; C024833:protein phosphatase inhibitor-2; D000097847:Protein Phosphatase Inhibitory Proteins