Enhancer sharing in a plasmid model containing the alpha-fetoprotein and albumin promoters.
It has been proposed that the alpha-fetoprotein (AFP) enhancers also regulate the adjacent albumin gene, since the -10 kbp albumin enhancer is inactive in a number of cell lines that express albumin. In transfection experiments, the AFP enhancers strongly stimulate the albumin promoter in cells that silence the AFP promoter. These observations led us to develop a plasmid model of AFP-albumin gene switching, in which the albumin and AFP promoters would compete for the three AFP enhancers. However, when AFPCAT + ALBgal genes were combined with the AFP enhancers in one plasmid, both genes were driven at full activity. There was no change in the relative promoter expression over a wide range of transfected DNA concentrations, demonstrating that relative promoter activity was independent of DNA concentration and of promoter concentration, and that neither promoter was limiting the expression of the other. In contrast, a control plasmid containing two albumin promoters showed mutual inhibition, indicating the expected promoter competition. The albumin and AFP promoters noncompetitively shared the three enhancers on this plasmid, resulting in high levels of transcription from both promoters.
Jin
J R
JR
Department of Pathology, School of Medicine, University of Pittsburgh, PA 15261.
Wen
P
P
Locker
J
J
D000418:Albumins / Q000235:genetics*; D000818:Animals; D004742:Enhancer Elements, Genetic; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D006801:Humans; D010957:Plasmids; D011401:Promoter Regions, Genetic; D051381:Rats; D014158:Transcription, Genetic; D014162:Transfection; D014407:Tumor Cells, Cultured; D000509:alpha-Fetoproteins / Q000235:genetics*
D000418:Albumins; D000509:alpha-Fetoproteins