title: Specific cellular proteins bind to critical promoter sequences of the adenovirus early EIIa promoter.
abstract: As an approach to the identification of essential factors required for specific expression of the adenovirus type 2 EIIaE early (EIIaE) promoter, an in vitro system was established. Under appropriate conditions, using crude extracts of non-infected HeLa cells, efficient and accurate EIIaE expression has been reproduced. As in vivo, this transcription was strongly dependent upon the integrity of two non-consensus TATA-like elements, T1 and T2, corresponding to the major (EIIaE1) and minor (EIIaE2) start sites, respectively, as well as upon intact upstream elements (A, between -40 and -50 and B, between -70 and -90) common to both overlapping promoters. The implication of specific DNA-binding proteins in the transcriptional effects mediated by these elements was demonstrated by DNAse I footprinting analyses. Both crude nuclear extracts and partially purified fractions confer specific protection against DNAse I digestion to the T1 and B promoter elements defined above, and to a far upstream region (element C, between -110 and -150), which has previously been identified as a weaker promoter element by in vivo transcriptional studies. Separation of the T1 recognition factor from those which bind to the upstream elements B and C by chromatographic fractionation of the extracts has also been achieved.
mesh_terms: D017456:Adenovirus Early Proteins; D000260:Adenoviruses, Human / Q000235:genetics* / Q000378:metabolism; D000957:Antigens, Viral, Tumor / Q000235:genetics*; D001483:Base Sequence; D004279:DNA, Viral / Q000235:genetics* / Q000378:metabolism; D005796:Genes; D005814:Genes, Viral; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D009698:Nucleoproteins / Q000302:isolation & purification / Q000378:metabolism*; D009856:Oncogene Proteins, Viral / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D014158:Transcription, Genetic
chemical_list: D017456:Adenovirus Early Proteins; D000957:Antigens, Viral, Tumor; D004279:DNA, Viral; D009698:Nucleoproteins; D009856:Oncogene Proteins, Viral