Uncoupling of hormone-dependence from chaperone-dependence in the L701H mutation of the androgen receptor.
The mechanisms underlying androgen receptor (AR)-mediated progression of prostate cancer following androgen ablation have yet to be fully determined. On this basis we screened naturally occurring mutants of human AR for hormone-independent activity using a yeast model system. An initial screen of 43 different mutants revealed that ARs having a Leu701His mutation (AR(L701H)) exhibited hormone-independent activation of a lacZ reporter gene. The AR(L701H) mutant bound dihydrotestosterone to a similar extent as did wild type AR, although its ability to be induced by hormone for transactivation was reduced substantially. Subsequent studies focused on the dependence of AR(L701H) on molecular chaperones for folding to the active state. We found that AR(L701H) was highly dependent on Hsp90 for its hormone-independent activation, suggesting that this chaperone functions in AR(L701H) folding. However, the mutant did not respond specifically to increased levels of FKBP52, suggesting that this chaperone functions at the hormone-dependent activation stage in the folding process. Further studies of AR(L701H) in PC3 cells suggested that this mutant is prohibited from hormone-independent transactivation in mammalian cells. However, basal expression of a reporter gene by AR(L701H) was not impaired by the presence of 17-allylamino-17-demethoxygeldanamycin as was wild type AR, suggesting differential interactions of these receptors with molecular chaperones in animal cells.
Robzyk
Kenneth
K
Department of Medicine and Program in Molecular Pharmacology and Chemistry, Memorial Sloan-Kettering Cancer Center, New York, NY 10021, USA.
Oen
Handy
H
Buchanan
Grant
G
Butler
Lisa M
LM
Tilley
Wayne D
WD
Mandal
Atin K
AK
Rosen
Neal
N
Caplan
Avrom J
AJ
D000728:Androgens / Q000378:metabolism*; D045744:Cell Line, Tumor; D018841:HSP90 Heat-Shock Proteins / Q000378:metabolism*; D006639:Histidine / Q000235:genetics*; D006801:Humans; D007930:Leucine / Q000235:genetics*; D008024:Ligands; D008297:Male; D050505:Mutant Proteins / Q000378:metabolism; D009154:Mutation / Q000235:genetics*; D011471:Prostatic Neoplasms / Q000378:metabolism / Q000473:pathology; D017434:Protein Structure, Tertiary; D011944:Receptors, Androgen / Q000737:chemistry / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D015533:Transcriptional Activation / Q000235:genetics; D015003:Yeasts
C410798:AR protein, human; D000728:Androgens; D018841:HSP90 Heat-Shock Proteins; D008024:Ligands; D050505:Mutant Proteins; D011944:Receptors, Androgen; D029701:Saccharomyces cerevisiae Proteins; D006639:Histidine; D007930:Leucine