Hyperphosphorylation of p53 induced by okadaic acid attenuates its transcriptional activation function.
The tumor suppressor and transcriptional factor p53 is a phosphorylated protein. Its phosphorylation states are regulated by several protein kinases and phosphatases. In this study, the wild-type p53 was transfected and expressed in chronic myelogenous leukemia K-562 cells. Incubation of the transfected cells with okadaic acid, an inhibitor of serine phosphatases 2A and 1, induced hyperphosphorylation of p53 protein. The treatment also increased the steady state level of p53 protein in the cells. However, the hyperphosphorylated p53 protein was less active in promoting transcription mediated by two p53-binding DNA elements, the ribosomal gene cluster and the p53 consensus DNA-binding sequence. Nevertheless, the decreased transcription activation was not due to decreased binding of p53 to these elements, as analyzed by mobility shift DNA-binding assays. In addition, the treatment did not induce a conformational change in p53, as assayed by two conformation-specific anti-p53 monoclonal antibodies, PAb240 and PAb1620. These results suggest that the phosphorylation induced by okadaic acid may selectively modulate the transcription activation function of p53. Consequently, phosphorylation may represent a mechanism of p53 inactivation in tumor cells that harbor the wild-type p53.
Zhang
W
W
Department of Hematology, University of Texas M.D. Anderson Cancer Center, Houston 77030.
McClain
C
C
Gau
J P
JP
Guo
X Y
XY
Deisseroth
A B
AB
D001483:Base Sequence; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics; D004988:Ethers, Cyclic / Q000494:pharmacology*; D016158:Genes, p53 / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000378:metabolism / Q000473:pathology; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D019319:Okadaic Acid; D010766:Phosphorylation / Q000187:drug effects; D011487:Protein Conformation; D014158:Transcription, Genetic / Q000187:drug effects* / Q000235:genetics; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*
D004268:DNA-Binding Proteins; D004988:Ethers, Cyclic; D016159:Tumor Suppressor Protein p53; D019319:Okadaic Acid; D004247:DNA; D008156:Luciferases