title: Regulation of the uteroferrin gene promoter in endometrial cells: interactions among estrogen, progesterone, and prolactin.
abstract: Expression of the gene for the porcine transplacental iron transport protein uteroferrin (UF) is largely restricted to the uterus, where it is differentially regulated by estrogen (E) and progesterone (P). To study the regulatory mechanisms subserving these effects, a 2-kilobase genomic fragment corresponding to -2005 to 48 nucleotides of the UF gene was ligated up-stream to the reporter gene chloramphenicol acetyltransferase (CAT). This construct (UF-CAT) was transiently transfected into rabbit endometrial (HRE-H9), mouse fibroblastic (AKR-2B), and human choriocarcinoma (JEG-3) cells. The basal gene promoter activity of UF-CAT was exhibited in H9 cells, but not in AKR-2B or JEG-3 cells. In contrast, a simian virus-40 early promoter (SV2) was functional in all three cell lines. The H9 cells were used to examine steroid regulation of the UF gene promoter. The CAT expression in H9 cells primed with E and PRL, but not with E or PRL alone, was stimulated by P. In contrast, basal activity of SV2 in these cells was unaffected by hormones, singly or in combination. To examine the basis for the E/PRL-dependent response to P, levels of P and E receptors in H9 cells were quantified. PRL and E plus PRL increased the number of high affinity sites for P, but had little effect on levels of high affinity sites for E in treated vs. untreated H9 cells. In vivo administration of PRL to cyclic gilts had no effect on levels of endometrial UF mRNA and secreted UF protein; however, E- plus PRL-treated gilts had higher (P less than 0.05) levels of endometrial UF mRNA and luminal UF than PRL-treated gilts. These results demonstrate in vitro functional activity of the UF gene promoter and associated 5' flanking region and suggest that sequences within this region may mediate tissue-specific and steroid hormone-regulated expression of the UF gene. Moreover, interactions among E, PRL, and P modulate UF gene expression in vivo and in vitro.
authors.affiliation: Animal Science Department, University of Florida, Gainesville 32611-0691.
authors.forename: A E
authors.initials: AE
authors.lastname: Fliss
authors.forename: F J
authors.initials: FJ
authors.lastname: Michel
authors.forename: C L
authors.initials: CL
authors.lastname: Chen
authors.forename: A
authors.initials: A
authors.lastname: Hofig
authors.forename: F W
authors.initials: FW
authors.lastname: Bazer
authors.forename: J Y
authors.initials: JY
authors.lastname: Chou
authors.forename: R C
authors.initials: RC
authors.lastname: Simmen
mesh_terms: D000135:Acid Phosphatase; D000818:Animals; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D004347:Drug Interactions; D004717:Endometrium / Q000378:metabolism*; D004958:Estradiol / Q000494:pharmacology*; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D007527:Isoenzymes; D008667:Metalloproteins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D010052:Ovariectomy; D011374:Progesterone / Q000494:pharmacology*; D011388:Prolactin / Q000494:pharmacology*; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000378:metabolism; D011817:Rabbits; D013552:Swine; D000071681:Tartrate-Resistant Acid Phosphatase; D014162:Transfection
chemical_list: D007527:Isoenzymes; D008667:Metalloproteins; D012333:RNA, Messenger; D011374:Progesterone; D004958:Estradiol; D011388:Prolactin; D015500:Chloramphenicol O-Acetyltransferase; D000135:Acid Phosphatase; D000071681:Tartrate-Resistant Acid Phosphatase