TPA can overcome the requirement for EIa and together act synergistically in stimulating expression of the adenovirus EIII promoter.
We have examined the control of gene expression from the adenovirus early region III (Ad-EIII) promoter, which contains two previously defined elements, the AP1 and ATF sites. We found that the AP1 element is capable of mediating activation by the adenovirus immediate early (EIa) gene products. Consistent with studies demonstrating that the AP1 site mediates signal transduction in response to 12-O-tetradecanoylphorbol 13-acetate (TPA) we have shown that TPA can activate Ad-EIII expression and overcome the requirement for EIa. Together TPA and EIa elicited a synergistic response in expression from the Ad-EIII promoter during both transient expression assays and viral infections. This synergistic effect required the AP1 element. An EIII promoter construct, in which sequences upstream of the TATA box had been replaced with four AP1 sites, was responsive to TPA and EIa and in combination promoted the synergistic effect. The analysis of specific factors involved in transcription from the Ad-EIII indicated that proteins recognizing the ATF and AP1 sites were important in expression from this promoter in vitro. Purification of protein factors that specifically stimulated EIII expression resulted in the isolation of a set of factors of the AP1 family. Affinity purified AP1 recognized and activated transcription through both the AP1 and ATF elements. In addition, a protein fraction was identified with DNA binding activity specific for the ATF element. This fraction was dependent on the ATF site for transcriptional activity.
Buckbinder
L
L
Department of Biochemistry, Robert Wood Johnson Medical School, University of Medicine and Dentistry of New Jersey, Piscataway 08854-5635.
Miralles
V J
VJ
Reinberg
D
D
D017456:Adenovirus Early Proteins; D000260:Adenoviruses, Human / Q000235:genetics*; D001483:Base Sequence; D002872:Chromosome Deletion; D004268:DNA-Binding Proteins / Q000235:genetics; D015967:Gene Expression Regulation, Viral / Q000187:drug effects*; D005814:Genes, Viral; D006367:HeLa Cells / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D015345:Oligonucleotide Probes; D009856:Oncogene Proteins, Viral / Q000235:genetics*; D010957:Plasmids; D011401:Promoter Regions, Genetic / Q000187:drug effects*; D013755:Tetradecanoylphorbol Acetate / Q000378:metabolism*; D014158:Transcription, Genetic; D014162:Transfection; D015678:Viral Structural Proteins / Q000235:genetics*
D017456:Adenovirus Early Proteins; D004268:DNA-Binding Proteins; D015345:Oligonucleotide Probes; D009856:Oncogene Proteins, Viral; D015678:Viral Structural Proteins; D013755:Tetradecanoylphorbol Acetate