The retinoid X receptor (RXR), a nuclear receptor that is activated by 9-cis-retinoic acid (9cRA), can regulate transcription as a homodimer or as a heterodimer with numerous other receptors. It was previously shown that, in the absence of ligand, RXR self-associates into homotetramers which are transcriptionally silent, and that ligand-binding induces dissociation of RXR tetramers into active species, dimers and monomers. Here, the implications of tetramer formation by RXR for the ability of the receptor to heterodimerize with the retinoic acid and the vitamin D receptors (RAR and VDR) were studied. In addition, the effects of cognate ligands for RXR and for RAR and VDR on formation of the respective heterodimers were examined. The data indicate that RXR subunits that are sequestered in tetramers were not available for interactions with RAR or VDR and, consequently, that in the absence of a RXR ligand, only a small fraction of this receptor became involved in heterodimers. RXR-selective ligands led to tetramer dissociation, but also inhibited the formation of heterodimers, directing a significant fraction of RXR into homodimers. Ligand binding by either heterodimerization partner significantly stabilized the respective heterodimer. Thus, maximal heterodimerization was observed in the presence of both 9cRA, acting to release active RXR species from tetramers, and the partner's cognate ligand, acting to overcome the inhibitory effect of 9cRA on heterodimer formation. These observations suggest that, by modulating protein-protein interactions within homo- and hetero-oligomers of RXR, cognate ligands control the relative distribution of potential RXR-containing complexes, thereby determining the transcriptional pathways that may be invoked under particular conditions in vivo.