title: Cooperation between the human estrogen receptor (ER) and MCF-7 cell-specific transcription factors elicits high activity of an estrogen-inducible enhancer from the trout ER gene promoter.
abstract: The human estrogen receptor (hER) is expressed in breast cancer MCF-7 cells and plays a major role in tumorigenic processes. In this report, we demonstrate that MCF-7-specific factors can cooperate with the hER to increase its transactivation activity. We previously demonstrated that the rainbow trout ER (rtER) gene is up-regulated by the rtER protein itself, through an enhancer that contains an imperfect estrogen-responsive element (FP1 area). By performing footprinting experiments, we have delineated two other regulatory regions (FP2 and FP3 areas) in the 0.2-kb enhancer. We show, by transient transfections, that hER poorly transactivates this enhancer in CHO-K1 and Ishikawa cells whereas, in MCF-7 cells, transcriptional activation occurs at a level about 20-fold higher than when the enhancer estrogen-responsive element (FP1) is the only regulatory region included in the reporter gene. These results indicate that areas other than FP1 are important regulatory sites of this enhancer. Site-directed mutagenesis demonstrated that the FP1 area is absolutely necessary for induction by estradiol as well as for basal activity of this enhancer in MCF-7 cells. Gel shift experiments showed that MCF-7 cells contain a factor that binds to the FP3 area and is poorly expressed in all other tested cell lines. As suggested by site-directed mutagenesis and deletion experiments, this FP3-binding protein may enhance the hER transactivation ability in MCF-7 cells. These data reinforce the idea that cell-specific transcription factors cooperate with steroid receptors to achieve maximal induction of hormone-responsive genes.
authors.affiliation: Laboratoire de Biologie Moléculaire, Centre Nationale de la Recherche Scientifique, Institut National de la Recherche Agronomique/Université de Rennes 1, France.
authors.forename: G
authors.initials: G
authors.lastname: Lazennec
authors.forename: L
authors.initials: L
authors.lastname: Kern
authors.forename: G
authors.initials: G
authors.lastname: Salbert
authors.forename: D
authors.initials: D
authors.lastname: Saligaut
authors.forename: Y
authors.initials: Y
authors.lastname: Valotaire
mesh_terms: D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D001943:Breast Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D016466:CHO Cells / Q000378:metabolism; D006224:Cricetinae; D018983:DNA Footprinting; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D003850:Deoxyribonuclease I / Q000378:metabolism; D004742:Enhancer Elements, Genetic / Q000187:drug effects*; D004958:Estradiol / Q000494:pharmacology; D006801:Humans; D008099:Liver / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D017686:Oncorhynchus mykiss / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011960:Receptors, Estrogen / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism* / Q000494:pharmacology; D014158:Transcription, Genetic; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D004268:DNA-Binding Proteins; D011960:Receptors, Estrogen; D014157:Transcription Factors; D004958:Estradiol; D003850:Deoxyribonuclease I