Summary
We estimate that E. coli RNA polymerase is able to form stable, rifampicin-resistant, pre-initiation complexes with Adenovirus 2 DNA at three to six binding sites. The number of RNA chains initiated from such complexes has been determined from the incorporation of γ-32P-ATP and-GTP at two rifampicin concentrations (7 μg/ml and 24 μg/ml) and after pre-incubation at either 25 or 37°C. The total number of RNA chains initiated ranges from 2.6 per Ad 2 DNA molecule at a rifampicin concentration of 24 μg/ml and pre-incubation temperature of 25°C, to 5.4 per Ad 2 DNA molecule at a rifampicin concentration of 7 μg/ml and pre-incubation temperature of 37°C. Efficient initiation with GTP occurs only after pre-incubation at 37°C whereas initiation with ATP is equally as efficient at either pre-incubation temperature. Promoters for initiation with ATP have been localized to the leftmost 58% of the Ad 2 DNA molecule, defined by the EcoR.RI restriction endonuclease fragment A; promoters for initiation with GTP are located on the remaining 42% of the Ad 2 DNA molecule. It is likely that on Adenovirus 2 DNA each RNA chain is initiated from a unique binding site which constitutes a separate promoter for E. coli RNA polymerase.
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Surzycki, S.J., Surzycki, J.A., DeLorbe, W. et al. In vitro transcription of adenovirus 2 DNA. Molec. Gen. Genet. 143, 177–184 (1976). https://doi.org/10.1007/BF00266920
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DOI: https://doi.org/10.1007/BF00266920