Intermolecular NH2-/carboxyl-terminal interactions in androgen receptor dimerization revealed by mutations that cause androgen insensitivity.
Structural alignment of the human androgen receptor dimer was investigated by introducing steroid binding domain mutations that cause partial or complete androgen insensitivity into fusion proteins containing the full-length androgen receptor or the steroid binding domain. Most of the mutants had unchanged apparent equilibrium androgen binding affinity and increased dissociation rates of [3H]methyltrienolone and required increased dihydrotestosterone concentrations for transcriptional activation. In a 2-hybrid protein interaction assay in mammalian cells, the steroid binding domain interacts with an NH2-terminal-DNA binding domain fragment and with the full-length androgen receptor at physiological androgen concentrations in a dose-dependent manner. However, mutations at Val-889 and Arg-752 disrupt the NH2-/carboxyl-terminal interaction when introduced into the steroid binding domain fragment but not when present in the full-length androgen receptor. The N-C bimolecular interaction reduces the dissociation rate of bound androgen and slows the degradation rate of the carboxyl-terminal steroid binding domain fragment. The results suggest that steroid binding domain residues Val-889 and Arg-752 are critical to the NH2-/carboxyl-terminal interaction and that an intermolecular N-C interaction occurs during receptor dimerization that results in an antiparallel arrangement of androgen receptor monomers.
Langley
E
E
Laboratories for Reproductive Biology, University of North Carolina, Chapel Hill, North Carolina 27599, USA.
Kemppainen
J A
JA
Wilson
E M
EM
D000728:Androgens / Q000378:metabolism*; D000818:Animals; D016466:CHO Cells; D019556:COS Cells; D006224:Cricetinae; D019281:Dimerization; D006801:Humans; D008297:Male; D016296:Mutagenesis; D011485:Protein Binding; D011944:Receptors, Androgen / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D000728:Androgens; D011944:Receptors, Androgen