Characterization of ligand binding, DNA binding and phosphorylation of progesterone receptor by two novel progesterone receptor antagonist ligands.
In order to gain a better understanding of the distinctive mechanisms of the various types of antiprogestins, we have characterized in vitro ligand binding, specific DNA binding and phosphorylation of progesterone receptor (PR) from T47D cells after treatment of cells with progestins (progesterone, R5020) and antiprogestins (RU486, ZK98299, Org 31806 and Org 31710). Treatment of the cells with R5020 or PR antagonists, with the exception of ZK98299, resulted in a quantitative upshift of PR-A and PR-B indicative of ligand/DNA-induced phosphorylation of PR. Treatment of cells with RU486, Org 31710 or Org 31806, but not R5020 or ZK98299 resulted in detectable PR-progesterone response element complexes (PR-PREc) as assessed by gel mobility shift assay. Although treatment of cells with ZK98299, a type I PR antagonist, did not induce phosphorylation, the antiprogestins, Org 31806 and Org 31710, in a manner identical to RU486, did. Our data suggest that Org 31806 and Org 31710 affect properties of PR from T47D cells that are similar to RU486.
Hurd
C
C
Department of Biological Sciences, Oakland University, Rochester 48309-4401, USA.
Underwood
B
B
Herman
M
M
Iwasaki
K
K
Kloosterboer
H J
HJ
Dinda
S
S
Moudgil
V K
VK
D001667:Binding, Competitive; D015153:Blotting, Western; D004247:DNA / Q000187:drug effects / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004963:Estrenes / Q000494:pharmacology; D005663:Furans / Q000494:pharmacology; D006067:Gonanes / Q000494:pharmacology; D006727:Hormone Antagonists / Q000494:pharmacology*; D006801:Humans; D008024:Ligands; D015735:Mifepristone / Q000494:pharmacology; D010766:Phosphorylation / Q000187:drug effects; D045167:Progesterone Congeners / Q000494:pharmacology; D011397:Promegestone / Q000494:pharmacology; D011980:Receptors, Progesterone / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D012967:Sodium Dodecyl Sulfate; D014407:Tumor Cells, Cultured
D004963:Estrenes; D005663:Furans; D006067:Gonanes; D006727:Hormone Antagonists; D008024:Ligands; D045167:Progesterone Congeners; D011980:Receptors, Progesterone; C076196:Org 31710; C076197:Org 31806; D015735:Mifepristone; D012967:Sodium Dodecyl Sulfate; D004247:DNA; D011397:Promegestone; C053238:onapristone