Journal article
Identification of Putative Androgen Receptor Interaction Protein Modules
Molecular & cellular proteomics, Vol.6(2), pp.252-271
02/2007
DOI: 10.1074/mcp.M600169-MCP200
PMID: 17052974
Abstract
We have developed a novel androgen receptor (AR) expression system in the 293 human embryonic kidney cell line that recapitulates AR biochemical activity as a steroid hormone receptor in prostate cancer cells. We used this system to identify putative AR-binding proteins in the cytosolic and nuclear compartments of mammalian cells using a large scale co-immunoprecipitation strategy coupled to quantitative mass spectrometry. For example, the heat shock 70 and 90 chaperones, which are known regulators of steroid hormone receptor, were identified as AR-binding proteins. AR purification enriched for proteins involved in RNA processing, protein transport, and cytoskeletal organization, suggesting a functional link between AR and these protein modules in mammalian cells. For example, AR purification in the nuclear compartment led to the specific enrichment of α-actinin-4, clathrin heavy chain, and serine-threonine protein kinase C δ. Short interfering RNA knockdown studies and co-transcriptional reporter assays revealed that clathrin heavy chain possessed co-activator activity during AR-mediated transcription, whereas α-actinin-4 and protein kinase C δ displayed both co-activator and co-repressor activity during AR-mediated transcription that was dependent upon their relative expression levels. Lastly immunohistochemical staining of prostate tissue showed that α-actinin-4 levels decreased in the nucleus of high grade cancerous prostate samples, suggesting its possible deregulation in advanced prostate cancers as previously observed in late stage metastatic breast cancers. Taken together, these findings suggest AR binds to specific protein modules in mammalian cells and that these protein modules may provide a molecular framework for interrogating AR function in normal and cancerous prostate epithelial cells.
Details
- Title: Subtitle
- Identification of Putative Androgen Receptor Interaction Protein Modules
- Creators
- Rohini Jasavala - University of California, DavisHarryl Martinez - University of California, DavisJaykumar Thumar - University of ConnecticutArmann Andaya - University of California, DavisAnne-Claude Gingras - Samuel Lunenfeld Research Institute, Toronto, Ontario M5G 1X5, CanadaJimmy K. Eng - Fred Hutchinson Cancer Research CenterRuedi Aebersold - Institute for Systems BiologyDavid K. Han - University of ConnecticutMichael E. Wright - University of California, Davis
- Resource Type
- Journal article
- Publication Details
- Molecular & cellular proteomics, Vol.6(2), pp.252-271
- DOI
- 10.1074/mcp.M600169-MCP200
- PMID
- 17052974
- NLM abbreviation
- Mol Cell Proteomics
- ISSN
- 1535-9476
- eISSN
- 1535-9484
- Publisher
- Elsevier Inc
- Language
- English
- Date published
- 02/2007
- Academic Unit
- Molecular Physiology and Biophysics
- Record Identifier
- 9984297501202771
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