Amplification of the transcriptional signal mediated by the tandem cAMP response elements of the glycoprotein hormone alpha-subunit gene occurs through several distinct mechanisms.
cAMP stimulates transcription of the human glycoprotein hormone alpha-subunit gene in choriocarcinoma cells. Combined treatment with phorbol esters potentiates this effect. Tandem cAMP response elements (CREs) in the proximal 5'-flanking sequence mediate the effect of cAMP. In this report, we show that the CREs can also mediate the synergistic effect of phorbol esters. In addition to serving as an inducible cis-acting element, the two CREs act synergistically to increase basal transcription. We now provide direct evidence via equilibrium binding studies that tandem CREs bind their trans-acting factors cooperatively. However, the level of cooperativity is insufficient to explain the high degree of transcriptional synergism, suggesting that another element in the alpha-subunit promoter may be required for complete synergism. In support of this hypothesis, we show that synergism is drastically reduced when the CREs are removed from the context of their native promoter and linked to a heterologous promoter. Thus, amplification of the transcriptional signal mediated by the tandem CREs occurs through at least three distinct mechanisms. First, at the level of signal transduction by convergence of the A- and C-kinase pathways; second, through homotropic interactions of trans-acting factors binding to tandem CREs; and finally through heterotropic interaction of the two CREs with another, as yet, undefined cis-acting element(s) in the human alpha-subunit promoter.
Andersen
B
B
Department of Pharmacology, School of Medicine, Case Western Reserve University, Cleveland, Ohio.
Kennedy
G C
GC
Hamernik
D L
DL
Bokar
J A
JA
Bohinski
R
R
Nilson
J H
JH
D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D001483:Base Sequence; D002822:Choriocarcinoma / Q000473:pathology; D000242:Cyclic AMP / Q000502:physiology*; D004268:DNA-Binding Proteins / Q000502:physiology; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D005796:Genes; D015292:Glycoprotein Hormones, alpha Subunit / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D011247:Pregnancy; D011401:Promoter Regions, Genetic; D011494:Protein Kinases / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012045:Regulatory Sequences, Nucleic Acid; D015398:Signal Transduction; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014157:Transcription Factors / Q000502:physiology; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000378:metabolism; D014594:Uterine Neoplasms / Q000473:pathology
D004268:DNA-Binding Proteins; D015292:Glycoprotein Hormones, alpha Subunit; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D015124:8-Bromo Cyclic Adenosine Monophosphate; D000242:Cyclic AMP; D011494:Protein Kinases; D013755:Tetradecanoylphorbol Acetate