title: Transcriptional repression of the α7 nicotinic acetylcholine receptor subunit gene (CHRNA7) by activating protein-2α (AP-2α).
abstract: The CHRNA7 gene, which encodes the α7 nicotinic acetylcholine receptor (α7*nAChR), has been implicated as a candidate gene in schizophrenia. Expression of the α7*nAChR mRNA and protein are reduced in multiple regions of post-mortem brain from patients diagnosed with schizophrenia. Transcriptional regulation may therefore be an important mechanism for the regulation of this gene. A 230-bp proximal promoter fragment, necessary for transcription in cultured neuroblastoma cells, was used to study a putative AP-2α binding site. Mutation of the site indicates that AP-2α plays a negative role in regulating CHRNA7 transcription. This was confirmed through knockdown and overexpression of AP-2α. Electrophoretic mobility shift assays (EMSAs) identified positive DNA-protein interaction at this same site, and supershift assays indicate that the complex includes AP-2α. The interaction was confirmed in cells using chromatin immunoprecipitation (ChIP). DNA methylation was discovered as an anomalous mechanism for CHRNA7 regulation in one cell line. These studies suggest a role for AP-2α regulation of CHRNA7 mRNA expression in multiple tissues during development.
authors: Departments of Biochemistry and Molecular Genetics, University of Colorado Denver Anschutz Medical Campus, Aurora, Colorado 80045; Department of Psychiatry, University of Colorado Denver Anschutz Medical Campus, Aurora, Colorado 80045.
authors: Jessica
authors: J
authors: Finlay-Schultz
authors: Department of Psychiatry, University of Colorado Denver Anschutz Medical Campus, Aurora, Colorado 80045.
authors: Andrew
authors: A
authors: Canastar
authors: Denver Veterans Affairs Medical Center, Denver, Colorado 80220.
authors: Margaret
authors: M
authors: Short
authors: Department of Psychiatry, University of Colorado Denver Anschutz Medical Campus, Aurora, Colorado 80045.
authors: Mohamed
authors: M
authors: El Gazzar
authors: Biological Sciences Department, University of Denver, Denver, Colorado 80208.
authors: Christina
authors: C
authors: Coughlan
authors: Department of Psychiatry, University of Colorado Denver Anschutz Medical Campus, Aurora, Colorado 80045; Denver Veterans Affairs Medical Center, Denver, Colorado 80220; Department of Pharmacology, University of Colorado Denver Anschutz Medical Campus, Aurora, Colorado 80045. Electronic address: Sherry.Leonard@ucdenver.edu.
authors: Sherry
authors: S
authors: Leonard
mesh_terms: D045744:Cell Line, Tumor; D019175:DNA Methylation; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D011978:Receptors, Nicotinic / Q000096:biosynthesis* / Q000378:metabolism; D012559:Schizophrenia / Q000235:genetics*; D050656:Transcription Factor AP-2 / Q000378:metabolism*; D014158:Transcription, Genetic; D064569:alpha7 Nicotinic Acetylcholine Receptor
chemical_list: C579167:Chrna7 protein, human; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D011978:Receptors, Nicotinic; D050656:Transcription Factor AP-2; D064569:alpha7 Nicotinic Acetylcholine Receptor