Promoter elements and transcriptional control of the mouse acetylcholinesterase gene.
The 5'-untranslated region of the mouse acetylcholinesterase gene has been characterized structurally by RNase protection, primer extension, and sequencing. Evidence has been obtained for the use of two alternative promoters in brain. Tissue-specific splicing to alternative acceptor sites in the 5'-untranslated exons occurs in brain, muscle, and erythropoietic cells. cis elements 5' of the cap site that is predominantly used in these tissues and cells have been analyzed by deletion analysis of promoter-reporter gene constructs and by site-specific mutagenesis. The cap site is found 107 base pairs (bp) 5' of the translation start site. This region is devoid of CAAT or TATA sequences; further in the 5' direction 50 and 70 bp are tandem Egr-1 sites. The putative promoter has been coupled to the open reading frame of a luciferase reporter gene. Deletion analysis shows that this region largely accounts for tissue-specific transcription seen upon transfection of neuronal and muscle cells. Mutagenesis of the Egr-1 sites results in a marked loss of reporter gene activity, further substantiating the importance of this region in the control of transcription. cis elements in the promoter differ from those found for the genes encoding the various subunits of the nicotinic acetylcholine receptor, and distinct differences in control of transcription are evident when the respective reporter genes are transfected into C2 muscle cells.
Li
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Department of Pharmacology, University of California, San Diego, La Jolla 92093-0636.
Camp
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Rachinsky
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TL
Bongiorno
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Taylor
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P
D000110:Acetylcholinesterase / Q000235:genetics*; D017398:Alternative Splicing; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D001921:Brain / Q000201:enzymology*; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D005091:Exons; D015971:Gene Expression Regulation, Enzymologic; D015698:Genomic Library; D006801:Humans; D007438:Introns; D004915:Leukemia, Erythroblastic, Acute; D051379:Mice; D008969:Molecular Sequence Data; D009132:Muscles; D009838:Oligodeoxyribonucleotides; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011978:Receptors, Nicotinic / Q000235:genetics; D015183:Restriction Mapping; D016385:TATA Box; D014158:Transcription, Genetic; D014162:Transfection; D014407:Tumor Cells, Cultured
D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D011978:Receptors, Nicotinic; D004247:DNA; D000110:Acetylcholinesterase