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Rapid Communication Isolation and Characterization of the Promoter of the Human GABA A Receptor α1 Subunit Gene
Author(s) -
Kang Inwha,
Lindquist David G.,
Kinane T. Bernard,
Ercolani Louis,
Pritchard Gary A.,
Miller Lawrence G.
Publication year - 1994
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1046/j.1471-4159.1994.62041643.x
Subject(s) - gabaa receptor , protein subunit , biology , gene , genetics , isolation (microbiology) , computational biology , receptor , microbiology and biotechnology , neuroscience , bioinformatics
The GABA A receptor, as assessed by ligand binding and chloride flux measurement in vivo and in vitro, is down‐regulated in response to chronic benzodiazepine exposure. The mRNA levels of the α1 and γ2 subunits of the receptor are also reduced. We have isolated the promoter of the gene encoding the α1 subunit of the GABA A receptor to elucidate the regulatory mechanism of its expression. A DNA segment 650 bp long has been Isolated that includes 151 bp of untranslated 5’end of the cDNA sequence and 500 bp of potential promoter‐enhancer region. The transcriptional activity of this DNA segment linked to the firefly luciferase gene showed a strong orientation specificity. The promoter activity was localized to a 60‐bp segment by deletion mapping. Mobility shift binding assay results suggest that this segment may interact with one or more factors in HeLa cell nuclear extracts to form a transcriptional complex. Primary cultures of embryonic chick cortical cells transfected with the promoter‐luciferase construct were treated chronically with lorazepam. Transcriptional activity of this promoter construct was strongly repressed by chronic administration of lorazepam.