The 3' flanking sequence of kappa promoter octamers was found to contain either a conserved A or G residue which increased the affinity of the octamer core motif for Octl and Oct2A. By transient transfections it was shown that decreasing the affinity of an octamer for Oct binding was crippling the transcription unit when the octamer was used in a minimal promoter, while it had only marginal effects when it was analysed in the context of an intact kappa promoter. As the octamer in a kappa promoter was replaced by a TAATGARAT motif with equal affinity for Oct protein binding the latter could still participate in synergistic transcriptional stimulation. Thus, the synergistic interactions involved in kappa promoter transcriptional stimulation are dependent on the presence of Oct proteins but not on the octamer DNA motif per se. Since the transcriptional coactivator OCA-B cannot interact with Oct protein bound to the TAATGARAT motif, the role of OCA-B in these interactions seems to be limited.