Estrogen receptor inducibility of the human Na+/H+ exchanger regulatory factor/ezrin-radixin-moesin binding protein 50 (NHE-RF/EBP50) gene involving multiple half-estrogen response elements.
The Na+/H+ exchanger regulatory factor (NHE-RF; also known as ezrin-radixin-moesin binding protein 50) is a primary response gene under estrogen receptor (ER) control that may provide a link between estrogen action and the regulation of cytoskeletal and cell-signaling pathways. These studies were undertaken to define the human NHE-RF genomic regions and regulatory sequences mediating its robust estrogen responsiveness. Screening of a human genomic library yielded NHE-RF clones comprising the full gene, including the 5'-regulatory region and first exon, which were found to contain a large number (13) of consensus half-estrogen response elements (EREs), but to lack palindromic full EREs. Transfection-transactivation assays with wild-type and mutant ERs and reporter gene constructs linked to progressive deletions, or containing mutations, of the 5'-flanking region including a portion of exon I, and electrophoretic mobility and competitive gel shift assays were performed. These demonstrated direct ER interaction with the multiple half-ERE sites and the importance of the one proximal half-ERE and the multiple upstream half-EREs for eliciting the robust transcription activation of the NHE-RF gene by the estrogen-ER complex. Our findings highlight a paradigm for gene regulation via numerous half-ERE sites that expands the range of modes by which DNA recognition sites mediate the actions of this nuclear receptor.
Ediger
Tracy R
TR
Department of Cell and Structural Biology, University of Illinois and College of Medicine, Urbana, Illinois 61801-3704, USA.
Park
Seong-Eun
SE
Katzenellenbogen
Benita S
BS
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D002352:Carrier Proteins / Q000235:genetics*; D002460:Cell Line; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D005091:Exons; D005809:Genes, Regulator; D006801:Humans; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D010750:Phosphoproteins / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011960:Receptors, Estrogen / Q000378:metabolism*; D017923:Sodium-Hydrogen Exchangers / Q000378:metabolism; D016329:Sp1 Transcription Factor / Q000378:metabolism
D002352:Carrier Proteins; D018076:DNA, Complementary; D010750:Phosphoproteins; D011960:Receptors, Estrogen; D017923:Sodium-Hydrogen Exchangers; D016329:Sp1 Transcription Factor; C093376:sodium-hydrogen exchanger regulatory factor