Peptide insertion in the DNA-binding domain of fish glucocorticoid receptor is encoded by an additional exon and confers particular functional properties.
The trout glucocorticoid receptor (rtGR) contains an additional sequence of nine amino acids located between the two zinc fingers of the DNA-binding domain (DBD) (Endocrinology 136 (1995) 3774). Polymerase chain reaction on trout genomic DNA and sequencing were performed in the DBD region, demonstrating that this peptide is encoded by an additional exon of 27 nucleotides between the two exons encoding the two zinc fingers of other nuclear receptors. This additional sequence in the rtGR confers a better binding affinity of the receptor to a single GRE, as shown by gel shift experiments with GST-DBDrtGR fusion proteins, deleted or not of the nine amino acids (Delta9). This higher affinity is correlated with a higher constitutive transcriptional activity of the receptor on a reporter gene driven by a single GRE, but not with the ligand-induced transcriptional activity. Nevertheless, on a double GRE, the wild type and rtGR-Delta9 are equally active on both constitutive or dexamethasone-induced transcriptional activity. This original DBD structure could have emerged during evolution such as to allow better regulation of glucocorticoid dependent genes in relation to the large spectrum of cortisol physiological functions in fish.
Lethimonier
C
C
Endocrinologie Moléculaire de la Reproduction, UMR-CNRS 6026 Bât 13, Campus de Beaulieu, Université de Rennes 1, 35042 Rennes cedex, France.
Tujague
M
M
Kern
L
L
Ducouret
B
B
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites / Q000235:genetics; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics; D003907:Dexamethasone / Q000494:pharmacology; D005091:Exons; D008969:Molecular Sequence Data; D010446:Peptide Fragments; D011485:Protein Binding / Q000235:genetics; D017434:Protein Structure, Tertiary; D011965:Receptors, Glucocorticoid / Q000737:chemistry / Q000235:genetics*; D020218:Response Elements; D015533:Transcriptional Activation; D014337:Trout
D004268:DNA-Binding Proteins; D010446:Peptide Fragments; D011965:Receptors, Glucocorticoid; D003907:Dexamethasone