Specific cellular proteins bind to critical promoter sequences of the adenovirus early EIIa promoter
| Autor: | Taka'aki Tamura, Charlotte Hauss, Hélène Boeuf, Deborah A. Zajchowski, Claude Kedinger |
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| Rok vydání: | 1987 |
| Předmět: |
Genes
Viral Transcription Genetic Plasma protein binding Biology DNA-binding protein Transcription (biology) In vivo Genetics Humans Antigens Viral Tumor Promoter Regions Genetic Gene Base Sequence Adenoviruses Human Adenovirus Early Proteins DNase-I Footprinting Promoter Oncogene Proteins Viral Molecular biology Cell biology Nucleoproteins Genes DNA Viral Deoxyribonuclease I HeLa Cells Protein Binding |
| Zdroj: | Nucleic Acids Research. 15:509-527 |
| ISSN: | 1362-4962 0305-1048 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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