Progesterone enhances target gene transcription by receptor free of heat shock proteins hsp90, hsp56, and hsp70.
Steroid receptors regulate transcription of target genes in vivo and in vitro in a steroid hormone-dependent manner. Unoccupied progesterone receptor exists in the low-salt homogenates of target cells as a functionally inactive 8 to 10S complex with several nonreceptor components such as two molecules of 90-kDa heat shock protein (hsp90), a 70-kDa heat shock protein (hsp70), and a 56-kDa heat shock protein (hsp56). Ligand-induced dissociation of receptor-associated proteins such as hsp90 has been proposed as the mechanism of receptor activation. Nevertheless, it has not been established whether, beyond release of heat shock proteins, the steroidal ligand plays a role in modulating receptor activity. To examine whether the release of these nonreceptor proteins from receptor complex results in a constitutively active receptor, we isolated an unliganded receptor form essentially free of hsp90, hsp70, and hsp56. Using a recently developed steroid hormone-responsive cell-free transcription system, we demonstrate for the first time that the dissociation of heat shock proteins is not sufficient to generate a functionally active receptor. This purified receptor still requires hormone for high-affinity binding to a progesterone response element and for efficient transcriptional activation of a target gene. When an antiprogestin, Ru486, is bound to the receptor, it fails to promote efficient transcription. We propose that in the cell, in addition to the release of receptor-associated inhibitory proteins, a distinct hormone-mediated activation event must precede efficient gene activation.
Bagchi
M K
MK
Department of Cell Biology, Baylor College of Medicine, Houston, Texas 77030.
Tsai
S Y
SY
Tsai
M J
MJ
O'Malley
B W
BW
D015153:Blotting, Western; D002474:Cell-Free System; D006367:HeLa Cells; D006360:Heat-Shock Proteins / Q000378:metabolism*; D006801:Humans; D015735:Mifepristone / Q000494:pharmacology; D011233:Precipitin Tests; D011374:Progesterone / Q000494:pharmacology*; D011980:Receptors, Progesterone / Q000737:chemistry / Q000187:drug effects / Q000302:isolation & purification / Q000502:physiology*; D014158:Transcription, Genetic / Q000187:drug effects*; D014407:Tumor Cells, Cultured
D006360:Heat-Shock Proteins; D011980:Receptors, Progesterone; D015735:Mifepristone; D011374:Progesterone