Transcriptional regulation of choline acetyltransferase gene by cyclic AMP.
The effect of cyclic AMP on the gene expression of choline acetyltransferase (ChAT) was studied in NG108-15, mouse neuroblastoma and rat glioma hybrid cell lines. Addition of dibutyryl cyclic AMP to the culture medium increased both the ChAT mRNA level and ChAT activity twofold. Polymerase chain reaction analysis of the ChAT mRNA indicated that, among the multiple mRNA species, M-type mRNA was transcribed most efficiently, with or without the addition of dibutyryl cyclic AMP. The 5' region of the mouse ChAT gene was ligated to the bacterial chloramphenicol acetyltransferase gene, and the expression of chloramphenicol acetyltransferase activity was determined by transfection analysis. Cyclic AMP derivatives enhanced the reporter gene expression in both transiently and stably transfected cells. DNA deletion analysis indicated that the intron region downstream of the M-type exon is necessary for the cyclic AMP responsiveness, and that cyclic AMP derivatives increase ChAT gene transcription mainly from M-type promoter. These results suggest that a cis-acting DNA element that confers the cyclic AMP responsiveness of the ChAT gene is present in the intron downstream of the M-type exon.
Department of Neurology, Tokyo Metropolitan Institute for Neuroscience, Japan.
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Misawa
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Takahashi
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Deguchi
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D003994:Bucladesine / Q000494:pharmacology; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D002795:Choline O-Acetyltransferase / Q000235:genetics*; D000242:Cyclic AMP / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects*; D005910:Glioma / Q000378:metabolism; D006822:Hybrid Cells; D051379:Mice; D008969:Molecular Sequence Data; D009447:Neuroblastoma / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011993:Recombinant Fusion Proteins; D014158:Transcription, Genetic; D014162:Transfection; D014407:Tumor Cells, Cultured
D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D003994:Bucladesine; D000242:Cyclic AMP; D015500:Chloramphenicol O-Acetyltransferase; D002795:Choline O-Acetyltransferase