Urea-induced transformation of native estrogen receptor and evidence for separate DNA-and ATP-binding sites.
We have investigated the involvement of hydrophobic receptor domains during transformation of the native estrogen receptor to a form(s) with high affinity for immobilized DNA and ATP. In the presence of 6 M urea the intact estrogen-receptor complex was completely (greater than 90%, n = 12) transformed into a DNA-binding configuration but only partially (35-45%, n = 8) transformed into an ATP-binding state. Similar experiments performed with unliganded receptor preparations further distinguished the receptor's DNA and ATP binding properties. While the urea-induced increase in receptor affinity for DNA-agarose was estrogen-dependent, the urea-induced increase in affinity for ATP-agarose was steroid-independent. This is the first direct evidence that hydrophobic receptor domains may be involved in the steroid-dependent exposure of the DNA binding site. This event is partially reversible and suggests that electrostatic interactions alone may not be sufficient to accurately describe receptor recognition of specific DNA acceptor sites.
D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D002417:Cattle; D002846:Chromatography, Affinity; D004247:DNA / Q000378:metabolism*; D004958:Estradiol / Q000378:metabolism; D005260:Female; D007700:Kinetics; D011960:Receptors, Estrogen / Q000378:metabolism*; D014508:Urea / Q000494:pharmacology*
D011960:Receptors, Estrogen; D004958:Estradiol; D000255:Adenosine Triphosphate; D014508:Urea; D004247:DNA