The intragenic enhancer of human immunodeficiency virus type 1 contains functional AP-1 binding sites.
An intragenic enhancer in the pol gene of human immunodeficiency virus type 1 has previously been identified (Verdin et al., Proc. Natl. Acad. Sci. USA 87:4874-4878, 1990). This element is composed of two subdomains both exhibiting phorbol ester-inducible enhancing activity on the viral thymidine kinase promoter in HeLa cells. Examination of the nucleotide sequence of one of these domains (nucleotides 4079 to 4342, HXB2 isolate) revealed the presence of three short DNA regions highly homologous to the recognition site for cellular transcription factor AP-1. Two short oligonucleotides containing these AP-1 sites each functioned as a phorbol ester-inducible enhancer when cloned upstream of the thymidine kinase promoter and transfected into HeLa cells. Gel mobility shift assays and competition experiments using the same two oligonucleotides demonstrated that they bound affinity-purified AP-1 or AP-1 present in uninduced and 12-O-tetradecanoylphorbol-13-acetate-induced HeLa nuclear extracts. Footprinting experiments confirmed that all three predicted sites bound purified AP-1. These results suggest that the AP-1 factor could play a role in the transcriptional regulation of human immunodeficiency virus type 1 gene expression.
Laboratory of Viral and Molecular Pathogenesis, National Institute of Neurological Disorders and Stroke, Bethesda, Maryland 20892.
C
C
Van Lint
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A
Burny
E
E
Verdin
D001483:Base Sequence; D001665:Binding Sites; D002467:Cell Nucleus / Q000187:drug effects / Q000502:physiology; D004279:DNA, Viral / Q000235:genetics* / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D005814:Genes, Viral; D015751:Genes, pol; D015497:HIV-1 / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D007438:Introns; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010957:Plasmids; D016755:Proto-Oncogene Proteins c-jun / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014158:Transcription, Genetic / Q000187:drug effects; D014162:Transfection
D004279:DNA, Viral; D009838:Oligodeoxyribonucleotides; D016755:Proto-Oncogene Proteins c-jun; D013755:Tetradecanoylphorbol Acetate