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Phosphorylation of androgen receptor isoforms
Hao Yun Wong1, Jan A Burghoorn, Marije Van Leeuwen
1Department of Reproduction and Development, Erasmus MC, P.O. Box 1738, 3000 DR, Rotterdam, The Netherlands.
The Biochemical Journal
|July 9, 2004
Summary
Androgen receptor (AR) isoforms arise from differential phosphorylation at specific sites. Mutating these phosphosites did not significantly alter AR
Area of Science:
- Molecular Biology
- Cellular Biology
- Biochemistry
Background:
- Phosphorylation of the human androgen receptor (AR) correlates with AR isoform appearance.
- The exact role of phosphorylation in AR isoform generation, specific phosphorylation sites, and their functions remain unclear.
Purpose of the Study:
- To investigate the extent of phosphorylation's involvement in AR isoform occurrence.
- To identify specific phosphorylation sites on the AR.
- To explore the functional significance of these phosphosites.
Main Methods:
- Expression of human AR in COS-1 cells.
- Mutational analyses (e.g., Ser-Ala substitutions).
- Reversed-phase High-Performance Liquid Chromatography (HPLC) and Mass Spectrometry (MS) for phosphorylation site identification.
Main Results:
- Three distinct AR isoforms (110 kDa, 112 kDa, 114 kDa) were identified.
- Ser-650 was constitutively phosphorylated; Ser-94 and Ser-515 were also implicated.
- Mutagenesis of identified phosphosites (S650A, S94A, S515A) generally did not affect AR's functional activity (transactivation, domain interactions).
- Ser-515 mutation modulated phosphorylation at Ser-650, suggesting cross-regulation.
Conclusions:
- AR isoform pattern is directly linked to differential phosphorylation of specific sites.
- While specific sites are involved in isoform generation, their mutation does not lead to significant changes in overall AR functional activity.