Functional cooperativity between two TPA responsive elements in undifferentiated F9 embryonic stem cells.
We have recently identified an enhancer, termed GPEI, in the 5'-flanking region of the rat glutathione transferase P gene, that is composed of two imperfect TPA (phorbol 12-O-tetradecanoate 13-acetate) responsive elements (TREs). Unlike other TRE-containing enhancers, GPEI exhibits a strong transcriptional enhancing activity in F9 embryonic stem cells. Mutational analyses have revealed that the high activity of GPEI is mediated by two imperfect TREs. Each TRE-like sequence has no activity by itself but acts synergistically to form a strong enhancer which is active even in the very low level of AP-1 activity in F9 cells. Furthermore, we show that synthetic DNAs containing two perfect TREs in certain arrangements have strong transcriptional enhancing activities in F9 cells and the activity is greatly influenced by the relative orientation and the distance of two TREs.
Okuda
A
A
Department of Biochemistry, University of Tokyo Faculty of Medicine, Japan.
Imagawa
M
M
Sakai
M
M
Muramatsu
M
M
D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation; D002460:Cell Line; D015342:DNA Probes; D004742:Enhancer Elements, Genetic / Q000187:drug effects*; D005982:Glutathione Transferase / Q000235:genetics*; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010957:Plasmids; D051381:Rats; D013724:Teratoma; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014162:Transfection
D015342:DNA Probes; D015345:Oligonucleotide Probes; D005982:Glutathione Transferase; D013755:Tetradecanoylphorbol Acetate