Androgen receptor phosphorylation - Regulation and identification of the phosphorylation sites.

被引:278
作者
Gioeli, D
Ficarro, SB
Kwiek, JJ
Aaronson, D
Hancock, M
Catling, AD
White, FM
Christian, RE
Settlage, RE
Shabanowitz, J
Hunt, DF
Weber, MJ [1 ]
机构
[1] Univ Virginia, Dept Microbiol, Hlth Sci Ctr, Charlottesville, VA 22908 USA
[2] Univ Virginia, Ctr Canc, Dept Pathol, Hlth Sci Ctr, Charlottesville, VA 22908 USA
[3] Univ Virginia, Ctr Canc, Dept Chem, Hlth Sci Ctr, Charlottesville, VA 22908 USA
[4] Univ Virginia, Ctr Canc, Dept Pharmacol, Ctr Hlth Sci, Charlottesville, VA 22908 USA
[5] Univ Virginia, Program Mol Med, Hlth Sci Ctr, Charlottesville, VA 22908 USA
关键词
D O I
10.1074/jbc.M204131200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Activation of signal transduction kinase cascades has been shown to alter androgen receptor (AR) activity. Although it has been suggested that changes in AR phosphorylation might be directly responsible, the basal and regulated phosphorylations of the AR have not been fully determined. We have identified the major sites of AR phosphorylation on ARs expressed in COS-1 cells using a combination of peptide mapping, Edman degradation, and mass spectrometry. We describe the identification of seven AR phosphorylation sites, show that the phosphopeptides seen with exogenously expressed ARs are highly similar to those seen with endogenous ARs in LN-CaP cells and show that specific agonists differentially regulate the phosphorylation state of endogenous ARs in LNCaP prostate cancer cells. Treatment of LNCaP cells with the synthetic androgen, R1881, elevates phosphorylation of serines 16, 81, 256, 308, 424, and 650. Ser-94 appears constitutively phosphorylated. Forskolin, epidermal growth factor, and phorbol 12-myristate 13-acetate increase the phosphorylation of Ser-650. The kinetics of phosphorylation of most sites in response to hormone or forskolin is temporally delayed, reaching a maximum at 2 h post-stimulation. The exception is Ser-81, which continues to display increasing phosphorylation at 6 h. These data provide a basis for analyzing mechanisms of crosstalk between growth factor signaling and androgen in prostate development, physiology, and cancer.
引用
收藏
页码:29304 / 29314
页数:11
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