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Cloning the uteroglobin gene promoter from the relic volcano rabbit ( Romerolagus diazi ) reveals an ancient estrogen‐response element
Author(s) -
AcostaMontesdeOca Adriana,
Zariñán Teresa,
Macías Héctor,
PérezSolís Marco A.,
UlloaAguirre Alfredo,
GutiérrezSagal Rubén
Publication year - 2012
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/mrd.22030
Subject(s) - biology , hormone response element , tata box , microbiology and biotechnology , promoter , uteroglobin , reporter gene , gene , transcription (linguistics) , luciferase , response element , mutant , estrogen receptor , genetics , gene expression , transfection , linguistics , philosophy , cancer , breast cancer
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. Mol. Reprod. Dev. 79: 337–345, 2012. © 2012 Wiley Periodicals, Inc.