DNA dependent recruitment of DDX17 and other interacting proteins by the human androgen receptor

Biochim Biophys Acta. 2009 Feb;1794(2):193-8. doi: 10.1016/j.bbapap.2008.11.001. Epub 2008 Nov 14.


An oligonucleotide-based assay (OBA) was used to identify novel co-factors that can be recruited by the deoxyribonucleic acid (DNA)-bound androgen receptor (AR). Nuclear extracts obtained from LNCaP cells, after incubation with R1881, were incubated with biotinylated oligonucleotides bound to streptavidin coated beads. The oligonucleotides contain 3 copies in tandem of the androgen responsive element ARE1 from the prostate specific antigen (PSA) gene promoter. As control incubation, a scrambled version of the tandem ARE1 was used. Immunoblots of the eluents revealed that the AR was bound to the ARE1 oligonucleotide and to a much lesser extent to the scrambled oligonucleotide. Proteins eluted from the oligonucleotides, were separated on a 5-15% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gradient gel, followed by identification using mass spectrometry. Identified proteins were scored for having one or more of the following known properties: nuclear localization, involved in transcription regulation, involvement in steroid hormone receptor (SHR) function, or specifical involvement in AR function. A total number of 85 nuclear proteins were found in two separate OBAs. Based on peptide counting, we found enrichment of 7 proteins eluted from the ARE1 oligonucleotide, compared to the scrambled oligonucleotide. Taken together with the obtained scores, these proteins are considered putative AR co-factors. One of these proteins, DDX17, is known to be a co-factor for estrogen receptor alpha (ERalpha), but has never been associated with AR function. The results indicate that the ARE oligonucleotide-based assay may allow enrichment of new candidate DNA-bound AR interacting proteins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biotin
  • Cell Line, Tumor
  • DEAD-box RNA Helicases / metabolism*
  • DNA / metabolism*
  • Humans
  • Male
  • Metribolone / pharmacology
  • Oligonucleotides
  • Promoter Regions, Genetic
  • Prostate-Specific Antigen / genetics
  • Protein Binding
  • Protein Interaction Mapping
  • Receptors, Androgen / genetics
  • Receptors, Androgen / physiology*
  • Response Elements
  • Streptavidin


  • Oligonucleotides
  • Receptors, Androgen
  • Metribolone
  • Biotin
  • DNA
  • Streptavidin
  • Prostate-Specific Antigen
  • DDX17 protein, human
  • DEAD-box RNA Helicases