THE Brn-3a POU family transcription factor activates a number of neuronal promoters which are repressed by the closely related Brn-3b factor. Although transcriptional activation by mutant forms of Brn-3a/Brn-3b can occur with a number of different amino acids at position 22 of the POU homeodomain, an isoleucine at this position is of critical importance for transcriptional repression. In addition, this isoleucine residue can mediate an interaction with the herpes simplex virus Vmw65 regulatory protein whereas the valine found in Brn-3a cannot do so. Thus, position 22 in the homeodomain plays a critical role in the function of neuronally expressed POU factors by controlling their interaction with viral and cellular co-activator/co-repressor proteins.