Sequence-specific DNA binding of the progesterone receptor to the uteroglobin gene: effects of hormone, antihormone and receptor phosphorylation.
The effects of ligand binding and receptor phosphorylation on the interaction of progesterone receptor with specific DNA sequences in the uteroglobin gene were studied by nitro-cellulose filter binding and DNase I footprinting. High affinity sites were mapped upstream from the transcription start and in the first intron. They contained a common TGTTCACT sequence. These sites were occupied with similar affinity by the receptor, either in its free state, or complexed with the hormone or an antagonist (RU486); and also by receptor which had been phosphorylated in vivo in a hormone-dependent manner. In all cases identical footprints were observed. These experiments led to the following conclusions. The hormone-dependency of receptor binding to DNA or chromatin is observed in intact cells and in crude cellular extracts but not with purified receptor. Thus in situ, the unliganded receptor probably interacts with some nuclear component(s) which stabilizes it in a 'non-activated' form (non-chromatin and non-DNA binding form). When isolated, the receptor may undergo activation, even in the absence of the hormone. Binding by receptor of an antihormone (and possibly receptor phosphorylation) exerts an effect on gene transcription through a mechanism which is different from (and probably follows) receptor interaction with the gene.
D000818:Animals; D001483:Base Sequence; D003600:Cytosol / Q000378:metabolism; D004247:DNA / Q000235:genetics* / Q000378:metabolism; D003850:Deoxyribonuclease I; D004963:Estrenes / Q000494:pharmacology; D005260:Female; D005796:Genes; D006023:Glycoproteins / Q000235:genetics*; D007700:Kinetics; D015735:Mifepristone; D010766:Phosphorylation; D011397:Promegestone / Q000494:pharmacology; D011817:Rabbits; D011980:Receptors, Progesterone / Q000187:drug effects / Q000378:metabolism*; D014598:Uteroglobin / Q000235:genetics*; D014599:Uterus / Q000378:metabolism
D004963:Estrenes; D006023:Glycoproteins; D011980:Receptors, Progesterone; D015735:Mifepristone; D004247:DNA; D014598:Uteroglobin; D011397:Promegestone; D003850:Deoxyribonuclease I