Actin cytoskeleton-dependent regulation of corticotropin-releasing factor receptor heteromers

Stress responses are highly nuanced and variable, but how this diversity is achieved by modulating receptor function is largely unknown. Corticotropin-releasing factor receptors (CRFRs), class B G protein-coupled receptors, are pivotal in mediating stress responses. Here we show that the two known CRFRs interact to form heteromeric complexes in HEK293 cells coexpressing both CRFRs and in vivo in mouse pancreas. Coimmunoprecipitation and mass spectrometry confirmed the presence of both CRF₁R and CRF₂βR, along with actin in these heteromeric complexes. Inhibition of actin filament polymerization prevented the transport of CRF₂βR to the cell surface but had no effect on CRF₁R. Transport of CRF₁R when coexpressed with CRF_2βR became actin dependent. Simultaneous stimulation of cells coexpressing CRF₁R+CRF₂βR with their respective high-affinity agonists, CRF+urocortin2, resulted in approximately twofold increases in peak Ca²⁺ responses, whereas stimulation with urocortin1 that binds both receptors with 10-fold higher affinity did not. The ability of CRFRs to form heteromeric complexes in association with regulatory proteins is one mechanism to achieve diverse and nuanced function.