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Nucleic Acids Research, 1989, Vol. 17, No. 17 6903-6914
© 1989

MOLECULAR BIOLOGY

Stereospecific relationships between elements in an SV40/Adenovirus-2 heterologous promoter

A.C. Lennard*, H.W.D. Matthes, J.M. Egly and P. Chambon

Laboratoire de Génétique Moléculaire des Eucaryotes du CNRS, Unité 184 de Biologie Moléculaire et de Génie Génétique de l'INSERM, Institut de Chimie Biologie, Faculté de Médecine 67085 Strasbourg, Cédex, France

Received June 12, 1989. Accepted August 4, 1989.

Spacing mutations have been constructed between an SV40 72 bp enhancer element and the Adenovirus-2 major late promoter upstream element (Ad2MLP-UE), the Ad2MLP-UE and the AdM2MLP TATA-box and the SV40 72 bp enhancer and the TATA-box in the absence of the AdM2MLP-UE. A clear periodic pattern of transcription from the Ad2MLP was obtained by inserting multiples of half of a DNA turn between the SV40 enhancer and the Ad2MLP-UE. We interpret this as a stereospecific requirement for the spatial alignment between the enhancer element and either the Ad2MLP-UE or the TATA-box. Insertions between the upstream element and the TATA-box did not give a periodic transcription pattern, though insertion of half of a DNA turn increased the steady-state level of specific RNA. Unexpectedly, spacing mutations between the enhancer and TATA-box in which the upstream element had been point mutated gave indications of stereospecificity, whereas the same insertions with a deleted upstream element showed little requirement for a precise stereoalignment.

*Present address ICRF, PO Box 123, Lincoln's Inn Fields, London WC2A 3PX, UK


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