Stereospecific relationships between elements in an SV40/adenovirus-2 heterologous promoter.
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 Ad2MLP TATA-box and the SV40 72 bp enhancer and the TATA-box in the absence of the Ad2MLP-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.
Lennard
A C
AC
Unité 184 de Biologie Moléculaire et de Génie Génétique de l'INSERM, Institut de Chimie Biologique, Faculté de Médecine, Strasbourg, France.
Matthes
H W
HW
Egly
J M
JM
Chambon
P
P
D000256:Adenoviridae / Q000235:genetics*; D004274:DNA, Recombinant; D004279:DNA, Viral / Q000235:genetics; D004742:Enhancer Elements, Genetic; D005814:Genes, Viral; D009690:Nucleic Acid Conformation; D011401:Promoter Regions, Genetic; D013539:Simian virus 40 / Q000235:genetics*; D014158:Transcription, Genetic
D004274:DNA, Recombinant; D004279:DNA, Viral