A simple technique for the prediction of interacting proteins reveals a direct Brn-3a-androgen receptor interaction

J Biol Chem. 2010 May 14;285(20):15286-15295. doi: 10.1074/jbc.M109.071456. Epub 2010 Mar 12.

Abstract

The formation of multiprotein complexes constitutes a key step in determining the function of any translated gene product. Thus, the elucidation of interacting partners for a protein of interest is of fundamental importance to cell biology. Here we describe a simple methodology for the prediction of novel interactors. We have applied this to the developmental transcription factor Brn-3a to predict and verify a novel interaction between Brn-3a and the androgen receptor (AR). We demonstrate that these transcription factors form complexes within the nucleus of ND7 neuroblastoma cells, while in vitro pull-down assays show direct association. As a functional consequence of the Brn-3a-AR interaction, the factors bind cooperatively to multiple elements within the promoter of the voltage-gated sodium channel, Nav1.7, leading to a synergistic increase in its expression. Thus, these data define AR as a direct Brn-3a interactor and verify a simple interacting protein prediction methodology that is likely to be useful for many other proteins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Blotting, Western
  • Cell Line
  • DNA Primers
  • Immunoprecipitation
  • Mice
  • Protein Binding
  • Rats
  • Receptors, Androgen / metabolism*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transcription Factor Brn-3A / metabolism*

Substances

  • DNA Primers
  • Receptors, Androgen
  • Transcription Factor Brn-3A