Tumor promoter phorbol ester reversibly modulates tyrosine dephosphorylatioin/ inactivation of protein kinase FA/GSK-3α in A431 cells
Corresponding Author
Shiaw-Der Yang
Institute of Biomedical Sciences, National Tsing Hua university, Taiwan, Republic of China
Institute of Basic Medicine, Chrng Gung Medical College, Tao-Yuan, Taiwan, Republic of China
Institute of Biomedical Sciences, National Tsing Hua University, Hsin-chu, Taiwan, Republic of ChinaSearch for more papers by this authorJau-Song Yu
Institute of Biomedical Sciences, National Tsing Hua university, Taiwan, Republic of China
Institute of Basic Medicine, Chrng Gung Medical College, Tao-Yuan, Taiwan, Republic of China
Search for more papers by this authorZin-Der Wen
Institute of Biomedical Sciences, National Tsing Hua university, Taiwan, Republic of China
Institute of Basic Medicine, Chrng Gung Medical College, Tao-Yuan, Taiwan, Republic of China
Search for more papers by this authorCorresponding Author
Shiaw-Der Yang
Institute of Biomedical Sciences, National Tsing Hua university, Taiwan, Republic of China
Institute of Basic Medicine, Chrng Gung Medical College, Tao-Yuan, Taiwan, Republic of China
Institute of Biomedical Sciences, National Tsing Hua University, Hsin-chu, Taiwan, Republic of ChinaSearch for more papers by this authorJau-Song Yu
Institute of Biomedical Sciences, National Tsing Hua university, Taiwan, Republic of China
Institute of Basic Medicine, Chrng Gung Medical College, Tao-Yuan, Taiwan, Republic of China
Search for more papers by this authorZin-Der Wen
Institute of Biomedical Sciences, National Tsing Hua university, Taiwan, Republic of China
Institute of Basic Medicine, Chrng Gung Medical College, Tao-Yuan, Taiwan, Republic of China
Search for more papers by this authorAbstract
The signal transducrion mechanism of protein kinase FA/GSK-3α by tyrosine phosphorylation in A431 cells was investigated. Kinase FA/GSK-3α was found to exist in a highly tyrosine-phosphorylated/activated state in resting cells but could be tyrosine-dephosphorylated and inactivated to ∼60% of the control level when cells were acutely treated with 1 μM tumor phorbol ester (TPA) at 37oC for 30 min, as demonstrated by metabolic 32P-labeling the cells, followed by immunoprecipitation and two-dimensional phosphoamino acid analysis and by immunodetection in an antikinase FA/GSK-3α immunoprecipitate kinase assay. Conversely, when cells were chronically treated with 1 μM TPA at 37°C for 24 h and processed under identical condetions, kinase FA/GSK-3α was found to be rephosphorylated on tyrosine residue and reactivated to ∼130% of the original control level. Taken together, the results provide initial evidence that the phosphotyrosine content and cellular activity of kinase FA/GSK-3α can be modulated in a reversible manner by short-term and long-term exposure of A431 cells to TPA. Since acute exposure of cells to TPA causes up-regulation of cellular protein kinase C (PKC) activity and prolonged exposure to TPA causes down-regulation of PKC, the results further suggest that the TPA-mediated modulation of PKC may play a role in the regulation of tyrosine phosphorylation and concurrent activation of kinase FA/GSK-3α in cells, representing a new mode of signal transduction pathway for the regulation of this multisubstrate/multifunctional protein kinase in cells.
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