title: Identification and characterization of a novel promoter of the mouse mu opioid receptor gene (Oprm) that generates eight splice variants.
abstract: Our previous studies isolated and characterized eight splicing variants containing exon 11 as the first coding exon. The location of exon 11 about 10 kb upstream from exon 1 dual promoters implied another promoter to drive expression of the exon 11 variants. I now identify the second promoter in the 5'-flanking region of exon 11. One major transcriptional start point was determined. Sequence analysis indicated that the 5'-flanking region of the exon 11 contained a putative TATA box, several CAAT boxes and a number of cis-acting elements. Functional analysis suggested that exon 11 promoter activity was most evident in neuronal-like cells. A basal core region containing the TATA box, a negative region and a positive region were identified. Electrophoretic mobility shift assays with the nuclear extracts from NIE-115 cells revealed several protein complexes that likely contained TATA box-associated factors, NF-1-like and cMyc-Max-like proteins, respectively. It also showed that a TATA-binding protein specifically bound to the TATA box fragment. Mutation analysis suggested that the TATA box in the basal core region played a fundamental role in the exon 11 promoter, whereas the NF-1 site acted as a positive element.
authors.affiliation: Laboratory of Molecular Neuro-Pharmacology, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, NY 10021, USA. pany@mskcc.org
authors.forename: Ying-Xian
authors.initials: YX
authors.lastname: Pan
mesh_terms: D024506:5' Flanking Region / Q000235:genetics; D017398:Alternative Splicing; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D016466:CHO Cells; D006224:Cricetinae; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D005091:Exons / Q000235:genetics; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D009687:Nuclear Proteins / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011485:Protein Binding; D017450:Receptors, Opioid, mu / Q000235:genetics*; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D016385:TATA Box / Q000235:genetics; D035181:TATA-Box Binding Protein / Q000378:metabolism; D024363:Transcription Initiation Site; D014407:Tumor Cells, Cultured
chemical_list: D009687:Nuclear Proteins; D017450:Receptors, Opioid, mu; D035181:TATA-Box Binding Protein; D004247:DNA