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.
Berwick
Daniel C
DC
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.
Diss
James K J
JKJ
Medical Molecular Biology Unit, University College London Institute of Child Health, 30 Guilford Street, London WC1N 1EH, United Kingdom.
Budhram-Mahadeo
Vishwanie S
VS
Medical Molecular Biology Unit, University College London Institute of Child Health, 30 Guilford Street, London WC1N 1EH, United Kingdom.
Latchman
David S
DS
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.
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