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Nucleic Acids Research, 1983, Vol. 11, No. 24 8735-8745
© 1983

MOLECULAR BIOLOGY

Far upstream sequences are required for efficient transcription from the adenovirus-2 E1A transcription unit

Paolo Sassone-Corsi, René Hen, Emiliana Borrelli, Todd Leff and Pierre Chambon*

Laboratoire de Génétique Moléculaire des Eucaryotes du CNRS Unité 184 de Biologic Moléculaire et de Génie Génétique de l'INSERM, Institut de Chimie Biologique, 11 rue Humann, 67085 Strasbourg Cédex, France

*To whom all reprint requests should be addressed

Received October 21, 1983. Revised November 24, 1983. Accepted November 24, 1983.

We have investigated the requirement for sequences located upstream from the TATA box for efficient transcription from the Adenovirus-2 (Ad2) E1A promoter. A series of deletions located within the E1A promoter upstream sequences were introduced into recombinants which contain or do not contain the E1A structural sequences. The amount of E1A-specific RNA produced after transfection into HeLa cells was determined by quantitative S1 nuclease analysis. We demonstrate that sequences located more than 231 bp upstream from the E1A capsite are required for efficient transcription from the E1A promoter. However, the requirement for these stimulatory sequences is less pronounced in recombinants which contain the E1A structural sequences than in those in which these sequences have been deleted. We demonstrate also that these Ad2 stimulatory sequences activate transcription in cis when inserted upstream from the heterologous –34 to +33 Ad2 major late promoter (Ad2MLP) element which is otherwise inactive when transfected into HeLa cells. These results suggest that the 270 bp Ad2 left-terminal segment contains an enhancer-like element.


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