Gene expressions regulated by N/C interacting and non-interactiong GR

Glucocorticoid receptor (GR) was demonstrated to undergo an N/C interaction between the 26VMDFY30 motif in the N-terminus and C-terminal LBD in the presence of its co-chaperone cytoplasmic constitutive active/androstane receptor retention protein (CCRP). Utilizing the fact that phosphor-mimic mutation of tyrosine 30 to glutamic acid abolished this N/C interaction, CCRP and either GR Y30F or GR Y30E mutant was ectopically expressed in COS-1 cells for subsequent cDNA microarray analysis. Dexamethasone, a potent GR ligand, significantly induced or repressed 673 and 689 genes in Y30F and Y30E, respectively. Among them, only 166 genes were commonly regulated. This analysis strongly suggested that the GR N/C interaction serves to regulate distinct gene sets. Gene expressions induced by dexametahsone treatment for 6 hours were measured in COS-1 cells were transiently transfected with mCCRP-V5 and FLAG-hGR (Y30F or Y30E). Three repricates were used in each group. As negative controls, samples prepared from 0.1% DMSO treated cell were used.