title: Cloning the uteroglobin gene promoter from the relic volcano rabbit (Romerolagus diazi) reveals an ancient estrogen-response element.
abstract: To gain further insight on the estrogen-dependent transcriptional regulation of the uteroglobin (UG) gene, we cloned the 5'-flanking region of the UG gene from the phylogenetically ancient volcano rabbit (Romerolagus diazi; Rd). The cloned region spans 812 base pairs (bp; -812/-1) and contains a noncanonical TATA box (TACA). The translation start site is 48 bp downstream from the putative transcription initiation site (AGA), and is preceded by a consensus Kozak box. Comparison of the Rd-UG gene with that previously isolated from rabbits (Oryctolagus cuniculus) showed 93% in sequence identity as well as a number of conserved cis-acting elements, including the estrogen-response element (ERE; -265/-251), which differs from the consensus by two nucleotides. In MCF-7 cells, 17β-estradiol (E(2)) induced transcription of a luciferase reporter driven by the Rd-UG promoter in a similar manner as in an equivalent rabbit UG reporter; the Rd-UG promoter was 30% more responsive to E(2) than the rabbit promoter. Mutagenesis studies on the Rd-ERE confirmed this cis-element as a target of E(2) as two luciferase mutant reporters of the Rd-promoter, one with the rabbit and the other with the consensus ERE, were more responsive to the hormone than the wild-type reporter. Gel shift and super-shift assays showed that estrogen receptor-α indeed binds to the imperfect palindromic sequence of the Rd-ERE.
authors.affiliation: Research Unit in Reproductive Medicine, Hospital de Gineco Obstetricia, No. 4 Dr. Luis Castelazo Ayala, Instituto Mexicano del Seguro Social, Tizapán San Angel, México, DF, Mexico.
authors.forename: Adriana
authors.initials: A
authors.lastname: Acosta-MontesdeOca
authors.forename: Teresa
authors.initials: T
authors.lastname: Zariñán
authors.forename: Héctor
authors.initials: H
authors.lastname: Macías
authors.forename: Marco A
authors.initials: MA
authors.lastname: Pérez-Solís
authors.forename: Alfredo
authors.initials: A
authors.lastname: Ulloa-Aguirre
authors.forename: Rubén
authors.initials: R
authors.lastname: Gutiérrez-Sagal
mesh_terms: D000818:Animals; D001483:Base Sequence; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D024202:Electrophoretic Mobility Shift Assay; D004958:Estradiol / Q000494:pharmacology; D047628:Estrogen Receptor alpha / Q000378:metabolism; D004967:Estrogens / Q000494:pharmacology / Q000502:physiology*; D005786:Gene Expression Regulation; D006801:Humans; D007793:Lagomorpha / Q000235:genetics*; D008969:Molecular Sequence Data; D010442:Peptide Chain Initiation, Translational; D011401:Promoter Regions, Genetic; D011817:Rabbits / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid; D020218:Response Elements; D016415:Sequence Alignment; D016385:TATA Box / Q000235:genetics; D024363:Transcription Initiation Site; D014158:Transcription, Genetic; D014598:Uteroglobin / Q000235:genetics*
chemical_list: D047628:Estrogen Receptor alpha; D004967:Estrogens; D004958:Estradiol; D014598:Uteroglobin