Reconstitution of ligand-mediated glucocorticoid receptor activity by trans-acting functional domains.
Glucocorticoid receptors (GRs) are ligand-inducible transcription factors that contain several functional domains. We tested whether GR activity can be reconstituted using domains expressed in separate molecules. Hence, we developed a general approach in which proteins can be individually expressed but interact specifically through the leucine zippers of c-Jun and c-Fos fused to each protein. The GR was divided into two different fragments, one encoding the N-terminal trans-activation and DNA-binding domains and conferring constitutive activity to a glucocorticoid-responsive reporter gene, and one containing the C-terminal, ligand-binding domain. Coexpression of the trans-activation-DNA-binding domain and the ligand-binding domain fragments leads to reconstituted ligand-regulated GR activity that is completely dependent on the presence of compatible zippers. These results suggest that, in GRs and perhaps other members of the steroid/thyroid hormone receptor superfamily, ligand-mediated function does not require that these domains be present in cis, but that they can also function in trans. This, together with the absence of interdomain dimerization signals, also suggests that these domains possibly evolved from separate genes.
Spanjaard
R A
RA
Department of Medicine, Brigham and Women's Hospital, Howard Hughes Medical Institute, Boston, Massachusetts 02115.
Chin
W W
WW
D000818:Animals; D005075:Biological Evolution; D004268:DNA-Binding Proteins / Q000378:metabolism; D005796:Genes; D016350:Leucine Zippers; D008024:Ligands; D017510:Protein Folding; D016760:Proto-Oncogene Proteins c-fos / Q000378:metabolism; D016755:Proto-Oncogene Proteins c-jun / Q000378:metabolism; D011965:Receptors, Glucocorticoid / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013329:Structure-Activity Relationship
D004268:DNA-Binding Proteins; D008024:Ligands; D016760:Proto-Oncogene Proteins c-fos; D016755:Proto-Oncogene Proteins c-jun; D011965:Receptors, Glucocorticoid; D011993:Recombinant Fusion Proteins