Dimerization of the human melanocortin 1 receptor: functional consequences and dominant-negative effects.

The melanocortin 1 receptor (MC1R), a G(S)-protein-coupled receptor (GPCR), is a key regulator of proliferation and differentiation of epidermal melanocytes, and a determinant of human skin phototype and cancer risk. Homodimerization has been demonstrated for several GPCRs, but little information is available for MC1R. SDS-PAGE analysis of melanoma cells and heterologous cells expressing epitope-tagged MC1R revealed dimeric and oligomeric species in detergent-solubilized extracts, confirmed by co-immunoprecipitation of differentially tagged MC1R forms. Dimerization occurs early during MC1R biosynthesis, and is seen for mutants displaying intracellular retention. These mutants exerted dominant-negative effects on wild-type (WT) MC1R. Conversely, partial functional trans-complementation of selected loss-of-function mutants was observed. WT-MC1R lacks cooperativity in agonist binding, yet coexpression of WT and a C-terminal deletion mutant yielded a form of different pharmacological properties. The natural diminished function alleles R151C, R160W, and D294H, associated with red hair, displayed dimerization and heterodimerization with WT. Coexpression of WT and R151C or R160W reduced the density of binding sites on the plasma membrane of transfected cells, whereas D294H mediated a dominant-negative effect on functional coupling to adenylyl cyclase. Therefore, subtle changes of functional properties may be associated with different MC1R haplotypes, contributing to the complexity of skin phenotype.