Estrogen and progesterone receptor-binding sites on the chicken vitellogenin II gene: synergism of steroid hormone action.
The chicken vitellogenin II gene is transcriptionally activated by estrogens. In transient transfection experiments in human T47D cells that contain receptors for various steroids, we showed estradiol, progestin, and androgen responses of a chimeric chicken vitellogenin II construct. This construct consists of DNA sequences from -626 to -590 upstream of the start of transcription of the chicken vitellogenin gene linked to the herpes simplex virus thymidine kinase promoter driving the transcription of the bacterial chloramphenicol acetyltransferase gene. Treatment of the transfected T47D cells with a combination of estradiol and the progestin R5020 led to a superinduction of chloramphenicol acetyltransferase activity, showing a synergistic action of these two steroids. This synergism was not observed upon treatment of the transfected cells with estradiol and the androgen dihydrotestosterone. Using point mutations in the vitellogenin gene fragment, we showed in functional and in in vitro DNase I footprinting assays with a purified progesterone receptor that, for the synergistic action of estradiol and R5020 to occur, the progesterone receptor must be bound to the vitellogenin gene fragment. The progesterone receptor-binding site was localized at -610 to -590, close to the consensus sequence (-626 to -613) for estrogen receptor binding and function. We therefore demonstrate here that two different steroid hormones can be functionally synergistic through the interaction of their corresponding receptors with two different binding sites adjacent to one another.
Cato
A C
AC
Kernforschungszentrum Karlsruhe, Institut für Genetik und Toxikologie, Federal Republic of Germany.
Heitlinger
E
E
Ponta
H
H
Klein-Hitpass
L
L
Ryffel
G U
GU
Bailly
A
A
Rauch
C
C
Milgrom
E
E
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002645:Chickens; D002678:Chimera; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D004357:Drug Synergism; D004958:Estradiol / Q000494:pharmacology*; D005786:Gene Expression Regulation; D005796:Genes; D006801:Humans; D008969:Molecular Sequence Data; D009649:Norpregnadienes / Q000494:pharmacology*; D011397:Promegestone / Q000494:pharmacology*; D011485:Protein Binding; D011960:Receptors, Estrogen / Q000378:metabolism*; D011980:Receptors, Progesterone / Q000378:metabolism*; D014158:Transcription, Genetic / Q000187:drug effects*; D014162:Transfection; D014819:Vitellogenins / Q000235:genetics*
D009649:Norpregnadienes; D011960:Receptors, Estrogen; D011980:Receptors, Progesterone; D014819:Vitellogenins; D004958:Estradiol; D011397:Promegestone; D015500:Chloramphenicol O-Acetyltransferase