Localization and hormonal stimulation of phosphorylation sites in the LNCaP-cell androgen receptor.
Phosphorylation of the androgen receptor in human prostate tumour cells (LNCaP) is increased by addition of androgens to intact cells. Double-label studies, using [35S]methionine incorporation into receptor protein, and [32P]P(i) to label metabolically receptor phosphorylation sites, have enabled us to determine the phosphate content, relative to receptor protein, of both nontransformed and transformed and androgen receptors generated in intact LNCaP cells. No net change in the phosphorylation of the intact 110 kDa steroid-binding component of the androgen-receptor complex was found upon transformation to the tight nuclear binding form in the intact cell. Partial proteolysis of androgen receptor protein metabolically labelled with [32P]P(i) and photolabelled with [3H]R1881 (methyltrienolone) revealed that phosphorylation occurs mainly in the N-terminal trans-activation domain, whereas no phosphorylation was detected in the steroid- and DNA-binding domains. The location of most (> 90%) of the hormonally regulated phosphorylation sites in the N-terminal trans-activation domain suggests a role of phosphorylation of the androgen receptor in transcription regulation.
Kuiper
G G
GG
Department of Endocrinology & Reproduction, Medical Faculty, Erasmus University Rotterdam, The Netherlands.
de Ruiter
P E
PE
Trapman
J
J
Boersma
W J
WJ
Grootegoed
J A
JA
Brinkmann
A O
AO
D000728:Androgens / Q000494:pharmacology*; D001665:Binding Sites; D002467:Cell Nucleus / Q000737:chemistry / Q000378:metabolism; D002918:Chymotrypsin / Q000378:metabolism; D003600:Cytosol / Q000737:chemistry; D004247:DNA / Q000378:metabolism; D006801:Humans; D007163:Immunosorbent Techniques; D008297:Male; D008715:Methionine / Q000378:metabolism; D015741:Metribolone / Q000378:metabolism; D010446:Peptide Fragments / Q000378:metabolism; D010710:Phosphates / Q000378:metabolism; D010766:Phosphorylation; D011471:Prostatic Neoplasms / Q000378:metabolism*; D011944:Receptors, Androgen / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D000728:Androgens; D010446:Peptide Fragments; D010710:Phosphates; D011944:Receptors, Androgen; D015741:Metribolone; D004247:DNA; D008715:Methionine; D002918:Chymotrypsin