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Nucleic Acids Research, 1985, Vol. 13, No. 11 4067-4083
© 1985

Articles

Cis and trans activation of adenovirus IVa2 gene transcription

Venkatachala Natarajan and Norman P. Salzman

Laboratory of Biology of Viruses, National Institute of Allergy and Infectious Diseases, National Institutes of Health Bethesda, MD 20205, USA

Received January 22, 1985. Revised May 9, 1985. Accepted May 10, 1985.

The transcriptional control region of the adenovnrus Iva2 promoter was analyzed by cloning this promoter in front of a gene coding for bacterial chloramphenicol acetyl transferase (CATase) and estimating levels of CATase and IVa2 promoter specific RNA synthesized after transfection. To produce detectable amounts of CATase with the IVa2 promoter, an enhancer has to be present in cis. In the absence of enhancer sequences, the adenovirus E1A gene can not stimulate CATase synthesis. When cells were transfected with plasmids containing enhancer sequences and various IVa2 mutant promoters upstream of the CAT gene, we observed that CATase activity was not reduced significantly even after deletion of all sequences upstream of the RNA initiation site

Synthesis of IVa2 specific RNA was dependent on plasmids containing an enhancer (SV40 72 bp repeat) that was present in cis. In the absence of enhancer sequences, co-transfection to provide the adenovirus E1A gene in trans also stimulated IVa2 RNA synthesis. When HeLa cells were transfected with various deletion mutants with an enhancer in cis it was seen that sequences -38 to -64 base pairs upstream of the RNA initiation site are necessary for efficient transcription. The E1A gene in trans and an enhancer in cis have an additive effect on RNA synthesis from both IVa2 and major late promoters

The basis for the conflicting results between transcription and CATase synthesis is discussed


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[Abstract] [Full Text] [PDF]


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