Identification of a region on the adenovirus E1A gene responsible for induction by phorbol ester tumor promoter.
12-O-Tetradecanoylphorbol-13-acetate (TPA) treatment induces human adenovirus (Ad) early region 1A (E1A) messenger ribonucleic acid expression in infected or Ad-transformed cells. Here, we report that deletion analysis has identified a TPA-responsive element (TRE) in the E1A enhancer region. Deletion analysis indicates that the TRE is located upstream of the E1A cap site between nucleotides -237 and -47. Incubation of extracts from TPA-treated cells with radioactively labeled deoxyribonucleic acid (DNA) fragments containing the TRE (-237 to -47) form specific DNA-protein complexes as demonstrated by gel shift analysis and Southwestern blotting. These experiments provide evidence that novel protein-DNA complexes are formed on a region of the E1A promoter required for TPA-enhanced expression. We speculate that these DNA-binding proteins may interact with the TRE and play a critical role in the mechanism through which TPA upregulates transcription from the Ad E1A gene.
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Department of Biomedical Sciences, Ohio University, Athens 45701, USA.
James
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D017500:Adenovirus E1A Proteins / Q000235:genetics*; D001665:Binding Sites; D026501:Blotting, Southwestern; D004247:DNA / Q000378:metabolism; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D004742:Enhancer Elements, Genetic; D017353:Gene Deletion; D015870:Gene Expression; D005786:Gene Expression Regulation / Q000187:drug effects; D006367:HeLa Cells; D006801:Humans; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000032:analysis; D020218:Response Elements; D015183:Restriction Mapping; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014162:Transfection
D017500:Adenovirus E1A Proteins; D012333:RNA, Messenger; D004247:DNA; D013755:Tetradecanoylphorbol Acetate; D004121:Dimethyl Sulfoxide