Hormone activation of baculovirus expressed progesterone receptors.
Human and chicken progesterone receptors (A form) were overproduced in a baculovirus expression system. These recombinant progesterone receptors were full-length bound progesterone specifically and were recognized by monoclonal antibodies, AB52 and PR22, specific for human and chicken progesterone receptor, respectively. In gel retardation studies, binding of recombinant human and chicken progesterone receptors to their progesterone response element (PRE) was specific and was enhanced in the presence of progesterone. Binding of human progesterone receptor to the PRE was also enhanced in the presence of the antiprogestin, RU486, but very little effect was observed in the presence of estradiol, dexamethasone, testosterone, and vitamin D. In our cell-free transcription system, human progesterone receptor induced transcription in a receptor-dependent and hormone-activable manner. Receptor-stimulated transcription required the presence of the PRE in the test template and could be specifically inhibited by excess PRE oligonucleotides. Furthermore, chicken progesterone receptor also induced in vitro transcription in a hormone-activable manner. These results demonstrate that steroid receptors overexpressed in a baculovirus expression system are functional and exhibit steroid-responsive binding and transcription. These observations support our present understanding of the mechanism of steroid receptor-regulated gene expression and provide a technological format for studies of the role of hormone and antihormone in altering gene expression.
Elliston
J F
JF
Department of Cell Biology, Baylor College of Medicine, Houston, Texas 77030.
Beekman
J M
JM
Tsai
S Y
SY
O'Malley
B W
BW
Tsai
M J
MJ
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D002460:Cell Line; D002645:Chickens; D003001:Cloning, Molecular; D005822:Genetic Vectors; D006801:Humans; D007313:Insecta; D007700:Kinetics; D011374:Progesterone / Q000378:metabolism / Q000494:pharmacology*; D016364:Reading Frames; D011980:Receptors, Progesterone / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic / Q000187:drug effects*; D014162:Transfection
D011980:Receptors, Progesterone; D011374:Progesterone