Mutations flanking the polyglutamine repeat in the modulatory domain of rat glucocorticoid receptor lead to an increase in affinity for hormone.

A polyglutamine repeat in the N-terminus of the rat glucocorticoid receptor shows polymorphism, with variants of Q2RQ5, Q2RQ15-21. We investigated whether these natural polymorphisms affect receptor function, and whether alleles with polyglutamine repeats shorter than Q2RQ5, between Q2RQ6-14, or longer than Q2RQ21 are not found naturally because they encode a dysfunctional receptor. Ligand binding and transactivation properties of sets of natural (Q2RQ5-Q2RQ21) and artificial (Q4-Q80) alleles were compared following expression in CV-1 cells. The sequence of artificial alleles at sites flanking the repeat region was altered slightly to facilitate cloning. Western blotting showed that all constructs expressed GR protein in CV-1 cells. When co-expressed with an MMTV-lacZ reporter plasmid, all GR proteins were shown to be transcriptionally active in the presence of hormone. Scatchard analysis of ligand binding curves showed that affinities for dexamethasone and corticosterone were not affected by variation in the polyglutamine repeat either the natural or artificial sets of alleles. However, affinities were greater for the artificial compared with the natural alleles (2-3-fold for dexametasone, p < 0.001; and 4-fold for corticosterone,p < 0.001). These differences provide evidence of a direct or indirect interaction within GR between the ligand binding domain and residues flanking the polyglutamine repeat of the N-terminal domain.