Glucocorticoid receptor lacking the tau 1 transactivation domain is a gene-specific regulator of the wild-type glucocorticoid-receptor activity.
The glucocorticoid receptor (GR) contains a major transactivation function (tau 1), located in the N-terminal domain. tau 1 contributes to about 80% of the ligand-inducible transcriptional activity of GR. In this study, we show that GR devoid of tau 1 (symbol: see text] GR) can inhibit activation of gene expression by wild-type GR but this does not occur for all target genes. Activation of the mouse mammary tumor virus promoter by wild-type GR in transiently transfected chinese hamster ovary (CHO) cells lacking endogenous GR was repressed by cotransfecting [symbol: see text] GR. This effect was proportional to the amount of transfected [symbol: see text] GR and was not due to squelching. A moderate expression level of stably transfected [symbol: see text] GR mutant was also shown to repress the transcriptional activity of endogenous GR present in rat skeletal myoblast L8 cells. Glucocorticoid mediated down regulation of endogenous GR gene expression can be blocked by the [symbol: see text] GR mutant in stably transfected L8 cells. In contrast, no inhibition was observed on glucocorticoid induction of the endogenous glutamine synthetase gene in L8 cells. However, glucocorticoid induction of a reporter gene driven by the chicken glutamine synthetase promoter was inhibited by [symbol: see text] GR in L8 cells. Stable expression of wild-type GR in CHO cells rendered the cells glucocorticoid responsive with regard to glutamine synthetase induction but coexpression of [symbol: see text] GR did not repress induction of the endogenous glutamine synthetase gene expression by wild-type GR. Expression of [symbol: see text] GR alone in CHO cells did not render the glutamine synthetase gene glucocorticoid responsive, indicating that [symbol: see text] GR has no transcriptional activity on the glutamine synthetase gene. We conclude from these results that the structure of glucocorticoid-response elements within target genes may be very critical for the ability of the mutant receptor to exhibit a dominant negative effect.
Delaunay
F
F
Department of Medical Nutrition, Karolinska Institute, Huddinge University Hospital, Novum, Sweden. frde@thon.esb.ki.se
Liden
J
J
Gustafsson
J A
JA
Okret
S
S
D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003907:Dexamethasone / Q000494:pharmacology; D015536:Down-Regulation / Q000187:drug effects; D005786:Gene Expression Regulation; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D005938:Glucocorticoids / Q000494:pharmacology; D005974:Glutamate-Ammonia Ligase / Q000235:genetics; D006801:Humans; D051381:Rats; D011965:Receptors, Glucocorticoid / Q000737:chemistry*; D017384:Sequence Deletion; D013329:Structure-Activity Relationship; D015533:Transcriptional Activation
D005938:Glucocorticoids; D011965:Receptors, Glucocorticoid; D003907:Dexamethasone; D005974:Glutamate-Ammonia Ligase