A 361 base pair region of the rat FSH-beta promoter contains multiple progesterone receptor-binding sequences and confers progesterone responsiveness.
The rat is frequently used as a model to study the role of progesterone (P) in regulating FSH secretion and synthesis. The ability of P to modulate rat FSH-beta mRNA levels suggests the presence of a functional hormone response element. We have found three PRE-like sequences upstream of the transcription start site in the rat FSH-beta gene. These sequences are herein referred to as PRE-like sequence #1, #2 and #3 with #1 being most distal from the start site. The current studies determined whether these PRE-like sequences bound P receptor (PR) and were functional in regulating the induction of expression by P. Electrophoretic mobility shift assays (EMSA) demonstrated that a single 289 base pair (bp) DNA fragment encompassing all three PRE-like sequences specifically bound PR. Further, PR bound with high affinity to double-stranded oligonucleotides representing individual PRE-like sequences #1, #2 and, with lower affinity to a double-stranded oligonucleotide representing PRE-like sequence, #3. We have cloned a 361 bp sequence from the promoter region of the rat FSH-beta gene encompassing all three PRE-like sequences into a luciferase reporter vector (pGL3-promoter) yielding pFSHbeta361-luc+ which when transiently transfected into primary rat pituitary cell cultures, conferred P-responsiveness to a heterologous promoter. P-responsiveness was dependent upon the presence of PR and was blocked by the PR antagonist RU-486. These data strongly suggest the presence of functional PRE's in the rat FSH-beta gene promoter.
O'Conner
J L
JL
Department of Physiology and Endocrinology, Medical College of Georgia, Augusta 30912-3000, USA. joconner@mail.mcg.edu
Wade
M F
MF
Prendergast
P
P
Edwards
D P
DP
Boonyaratanakornkit
V
V
Mahesh
V B
VB
D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004247:DNA / Q000378:metabolism; D005260:Female; D005640:Follicle Stimulating Hormone / Q000235:genetics*; D037201:Follicle Stimulating Hormone, beta Subunit; D005786:Gene Expression Regulation / Q000235:genetics*; D006609:High Mobility Group Proteins / Q000494:pharmacology; D006801:Humans; D015735:Mifepristone / Q000494:pharmacology; D008969:Molecular Sequence Data; D010903:Pituitary Gland, Anterior / Q000166:cytology; D011374:Progesterone / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D045167:Progesterone Congeners / Q000494:pharmacology; D011397:Promegestone / Q000494:pharmacology; D011401:Promoter Regions, Genetic / Q000235:genetics*; D051381:Rats; D011980:Receptors, Progesterone / Q000378:metabolism*; D011993:Recombinant Fusion Proteins; D014162:Transfection
D037201:Follicle Stimulating Hormone, beta Subunit; D006609:High Mobility Group Proteins; D045167:Progesterone Congeners; D011980:Receptors, Progesterone; D011993:Recombinant Fusion Proteins; D015735:Mifepristone; D011374:Progesterone; D005640:Follicle Stimulating Hormone; D004247:DNA; D011397:Promegestone