Biphasic activation of two mitogen-activated protein kinases during the cell cycle in mammalian cells

• Published in: The Journal of biological chemistry Vol. 267; no. 28; pp. 20293 - 20297
• Main Authors: TAMEMOTO, H, KADOWAKI, T, TOBE, K, UEKI, K, IZUMI, T, CHATANI, Y, KOHNO, M, KASUGA, M, YAZAKI, Y, AKANUMA, Y
• Format: Journal Article
• Language: English
• Published: Bethesda, MD American Society for Biochemistry and Molecular Biology 05.10.1992
• Subjects: activation; Amino Acid Sequence; Analytical, structural and metabolic biochemistry; Animals; antibodies; Biological and medical sciences; Blotting, Western; Calcium-Calmodulin-Dependent Protein Kinases - metabolism; CDC2 Protein Kinase - metabolism; cell cycle; Cell Cycle - drug effects; CHO Cells; Cricetinae; Enzyme Activation; Enzymes and enzyme inhibitors; Fundamental and applied biological sciences. Psychology; MAP kinase; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Mitogen-Activated Protein Kinases; Molecular Sequence Data; Nocodazole - pharmacology; Phosphorylation; Precipitin Tests; Protein-Tyrosine Kinases - metabolism; Threonine - metabolism; Thymidine - metabolism; Transferases; Tyrosine - metabolism; Vertebrata; Immunoprecipitation reaction; Phosphorylation; Mammalia; Enzymatic activity; Structure activity relation; Rodentia; Cell cycle; Hamster; CHO cell line
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/s0021-9258(19)88700-8

We studied mitogen-activated protein kinase (MAPK) activities during the cell cycle of Chinese hamster ovary (CHO) cells using site-specific antibodies against extracellular signal-regulated kinase-1, a 44-kDa MAPK (Boulton, T.G., Yancopoulos, G.D., Gregory, J.S., Slauer, C., Moomaw, C., Hsu, J., and Cobb, M.H. (1990) Science 249, 64-67). These antibodies detected two distinct MAPKs (44- and 42-kDa MAPKs) in CHO cells. CHO cells were arrested at metaphase in the M phase by treatment with nocodazole, and activities of MAPKs were analyzed at specific time points after release from arrest. Immune complex kinase assay and renaturation and phosphorylation assay in substrate-containing gel revealed that both 44- and 42-kDa MAPKs had activities in the G1 through S and G2/M phases and were activated biphasically, in the G1 phase and around the M phase. MAPKs were inactivated in metaphase-arrested cells. The amount of MAPKs did not change significantly in the cell cycle. In the G1, S, and G2/M phases, MAPKs were phosphorylated on both tyrosine and threonine residues and dephosphorylated in metaphase-arrested cells. Our data suggest that MAPKs may play some role in the cell cycle other than G0/G1 transition.