Control of transcription initiation in vitro requires binding of a transcription factor to the distal promoter of the ovalbumin gene.
We used a cell-free HeLa cell transcription system to identify and characterize transcription factors and the promoter elements that they recognize in RNA polymerase II-transcribed genes. Deletion of the region (-71 to -83) containing the GTCAAA direct repeat resulted in a marked decrease of specific transcription of the ovalbumin gene; transcription could be competed with DNA fragments containing this sequence. Furthermore, DNase I footprinting identified a protein-binding site including this direct repeat with crude extracts and one of the partially purified protein fractions required for transcription. We propose that a soluble factor activates transcription through binding to the direct repeat of GTCAAA sequence upstream from the ovalbumin gene.
D001483:Base Sequence; D002872:Chromosome Deletion; D003850:Deoxyribonuclease I / Q000378:metabolism; D006367:HeLa Cells; D066298:In Vitro Techniques; D010047:Ovalbumin / Q000235:genetics*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012319:RNA Polymerase II / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic
D014157:Transcription Factors; D010047:Ovalbumin; D012319:RNA Polymerase II; D003850:Deoxyribonuclease I