Translation-independent association of mRNAs encoding protomers of the 5-HT _2A -mGlu2 receptor complex in living cells

The serotonin 2A receptor (5-HT _2A R) and the metabotropic glutamate 2 receptor (mGluR2) form heteromeric G protein-coupled receptor (GPCR) complexes through a direct physical interaction. Co-translational association of mRNAs encoding subunits of heteromeric ion channels has been reported, but whether complex assembly of GPCRs occurs during translation remains unknown. Our in vitro data reveal evidence of co-translational modulation in 5-HT _2A R and mGluR2 mRNAs following siRNA-mediated knockdown. Interestingly, immunoprecipitation of either 5-HT _2A R or mGluR2, using an antibody targeting epitope tags at their N-terminus, results in detection of both transcripts associated with ribonucleoprotein complexes containing RPS24. Additionally, we demonstrate that the mRNA transcripts of 5-HT _2A R and mGluR2 associate autonomously of their respective encoded proteins. Validation of this translation-independent association is extended ex vivo using mouse frontal cortex samples. Together, these findings provide mechanistic insights into the co-translational assembly of GPCR heteromeric complexes, unraveling regulatory processes governing protein-protein interactions and complex formation.