A cis-acting sequence, located at -450 in the promoter of the human interferon-inducible gene 6-16, binds constitutively to a nuclear protein and decreases the expression of a reporter interferon-inducible promoter.
An interferon-inducible synthetic promoter was constructed with oligonucleotides which correspond to two regions of the promoter of the human interferon-inducible gene 6-16 that interact in-vitro with nuclear proteins. Firstly, we cloned a direct repeat sequence that was necessary for interferon regulation and that interacted with a 55 kilodalton nuclear protein. 5' to the direct repeat we introduced a 45 bp long oligonucleotide located at -450 in the native 6-16 promoter that interacted in-vitro with an 80 kilodalton nuclear protein. Expression after transfection of these plasmids showed that the direct repeat is necessary for interferon-inducible control and the -450 oligonucleotide by itself has no effect on transcriptional activity while in conjunction with the direct repeat it decreased the basal and interferon-inducible transcriptional activity of the reporter promoter in human cells.
Department of Immunology, University of Texas M.D. Anderson Cancer Center, Houston.
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Chernajovsky
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Kirby-Sanders
D000818:Animals; D001483:Base Sequence; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D004247:DNA / Q000378:metabolism; D005786:Gene Expression Regulation / Q000502:physiology; D006801:Humans; D007372:Interferons / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009687:Nuclear Proteins / Q000378:metabolism; D009841:Oligonucleotides / Q000378:metabolism; D010957:Plasmids; D011401:Promoter Regions, Genetic / Q000502:physiology*; D011485:Protein Binding; D014158:Transcription, Genetic
D009687:Nuclear Proteins; D009841:Oligonucleotides; D004247:DNA; D007372:Interferons; D015500:Chloramphenicol O-Acetyltransferase