A simple technique for the prediction of interacting proteins reveals a direct Brn-3a-androgen receptor interaction.
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.
Medical Molecular Biology Unit, University College London Institute of Child Health, 30 Guilford Street, London WC1N 1EH, United Kingdom. Electronic address: daniel.berwick@live.pharmacy.ac.uk.
Daniel C
DC
Berwick
Medical Molecular Biology Unit, University College London Institute of Child Health, 30 Guilford Street, London WC1N 1EH, United Kingdom.
James K J
JKJ
Diss
Medical Molecular Biology Unit, University College London Institute of Child Health, 30 Guilford Street, London WC1N 1EH, United Kingdom.
Vishwanie S
VS
Budhram-Mahadeo
Medical Molecular Biology Unit, University College London Institute of Child Health, 30 Guilford Street, London WC1N 1EH, United Kingdom; Birkbeck, University of London, Malet Street, London WC1E 7HX, United Kingdom.
David S
DS
Latchman
D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002460:Cell Line; D017931:DNA Primers; D047468:Immunoprecipitation; D051379:Mice; D011485:Protein Binding; D051381:Rats; D011944:Receptors, Androgen / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D050819:Transcription Factor Brn-3A / Q000378:metabolism*
D017931:DNA Primers; D011944:Receptors, Androgen; D050819:Transcription Factor Brn-3A