Sensitivity of progesterone receptors to aurintricarboxylic acid. Inhibition of nuclear uptake, ATP and DNA binding.
When hen oviduct cytosol samples containing progesterone receptor complexed to [3H]progesterone were incubated with isolated nuclei in presence of 0.2 mM aurintricarboxylic acid, more than 50% inhibition occurred in the uptake of progesterone receptor by the nuclei. The activated form of progesterone receptor appeared to be more sensitive to the presence of aurintricarboxylic acid since pretreatment of non-activated progesterone receptor with the inhibitor and the subsequent removal of the latter prior to activation did not result in the inhibition of receptor uptake by the nuclei. Also, the binding of progesterone receptor to columns of DNA-cellulose of ATP-Sepharose was abolished under similar conditions. When nuclei, ATP-Sepharose or DNA-cellulose were preincubated with the inhibitor prior to the addition of receptor preparations, no such inhibition resulted indicating that the inhibitor may be interacting with the receptor protein and not complexing to ATP, DNA or sites in the nuclei. The steroid binding properties of progesterone receptor, however, remained intact under these conditions. Both A and B forms of progesterone receptor are equally sensitive to aurintricarboxylic acid presence when tested for their nuclear uptake. Aurintricarboxylic acid was also found to be very effective at low concentrations (0.25 mM) in eluting the receptor complexes off ATP-Sepharose columns without disrupting the steroid binding properties of progesterone receptor. Our results suggest that aurintricarboxylic acid is an effective inhibitor of progesterone receptor and that it may be acting by interfering with a site(s) on progesterone receptor which may be exposed upon activation and are involved in such processes as ATP binding, nuclear uptake and DNA binding. These observations suggest that use of aurintricarboxylic acid as a chemical probe for the analysis of progesterone receptor.
D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D001312:Aurintricarboxylic Acid / Q000494:pharmacology*; D002467:Cell Nucleus / Q000378:metabolism; D002645:Chickens; D002846:Chromatography, Affinity; D003509:Cyclohexanecarboxylic Acids / Q000494:pharmacology*; D003600:Cytosol / Q000187:drug effects; D004247:DNA / Q000378:metabolism*; D005260:Female; D008660:Metabolism; D010057:Oviducts / Q000187:drug effects; D011374:Progesterone / Q000378:metabolism; D011980:Receptors, Progesterone / Q000187:drug effects*
D003509:Cyclohexanecarboxylic Acids; D011980:Receptors, Progesterone; D001312:Aurintricarboxylic Acid; D011374:Progesterone; D000255:Adenosine Triphosphate; D004247:DNA