Steroid controlled expression of the chicken lysozyme and the rat tryptophan oxygenase gene after transfer into eukaryotic cells.
To study the mechanism of steroid induced transcriptional control we have introduced recombinants of the chicken lysozyme gene and the rat tryptophan oxygenase (TO) gene into heterologous and homologous cells. To monitor the activity of the TO-promoter, 1.9 kb of the TO 5'-flanking sequences were fused with sequences coding for the bacterial enzyme chloramphenicol acetyltransferase (CAT). Upon transfer into mouse L-cells the transient expression of the TO-CAT recombinant was found to be inducible by dexamethasone. Transient expression of chicken lysozyme gene recombinants after introduction into various cell types could only be detected in chicken oviduct cells, or in conjunction with SV-40 enhancer sequences in human cells. The recombinant gene used in oviduct cells was a fusion between the lysozyme promoter, including 1.4 kb of upstream sequences, and the coding region of the gene for SV 40 T-antigen (plys-T). The expression in oviduct cells was stimulated by dexamethasone or progesterone, whereas SV 40 enhanced expression of lysozyme sequences in human cells could not be regulated by steroids. Using several deletion mutants, a region between -220 bp and -140 bp upstream of the cap site was found to be essential for both regulation by glucocorticoids as well as by progesterone.
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002645:Chickens; D004274:DNA, Recombinant / Q000378:metabolism*; D003907:Dexamethasone / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D051379:Mice; D008845:Microinjections; D009113:Muramidase / Q000235:genetics*; D010057:Oviducts / Q000378:metabolism; D011374:Progesterone / Q000494:pharmacology; D051381:Rats; D014158:Transcription, Genetic / Q000187:drug effects; D014366:Tryptophan Oxygenase / Q000235:genetics*
D004274:DNA, Recombinant; D011374:Progesterone; D003907:Dexamethasone; D014366:Tryptophan Oxygenase; D009113:Muramidase