Enhanced phosphorylation of progesterone receptor by protein kinase C in human breast cancer cells.
Affinity-isolated progesterone receptor (PR) from human breast cancer cells incubated with [32P]orthophosphate was shown to exist as a phosphoprotein. Exposure of the cells to 10 nM phorbol-12-myristate-13-acetate (PMA) for 10 min increased by 30-40% the amount of label incorporated into the 116-kDa receptor protein. A two-fold increase in the total number of steroid binding sites was also observed in cells receiving PMA treatment. This apparent unmasking of PR binding sites by phosphorylation probably involved conformational changes to existing receptor complexes and affected the eventual state of receptor dissociation or transformation. An increase primarily in the 8 S sedimenting molecular species was observed but PMA treatment also led to the appearance of a smaller, 2-3 S form of receptor (10% of total) that was not present in control samples. When cytosols were partially transformed in vitro by ATP and salt, all molecular species of receptor (8, 4, and 2-3 S) from the PMA-treated samples consistently migrated faster in sucrose gradients. The larger amount of 2-3 S receptor in PMA-treated samples disappeared when ATP, but not salt, was the transforming agent. These results suggest a major role for phosphorylating reactions in the receptor-mediated action of steroids by regulating hormone-binding and influencing receptor transformation. Tumor promoters such as the phorbol esters may act by artificially increasing the level of processing of steroid receptor.
Boyle
D M
DM
Department of Chemical Pathology, South African Institute for Medical Research, Johannesburg.
van der Walt
L A
LA
D001943:Breast Neoplasms / Q000378:metabolism*; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D008970:Molecular Weight; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism*; D011980:Receptors, Progesterone / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D011980:Receptors, Progesterone; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate