The primary goal was to test the hypothesis that agonist-induced CRF₁ receptor phosphorylation is required for arrestins to translocate from cytosol to cell membrane. We also sought to determine the relative importance to arrestin recruitment of motifs in the CRF₁ receptor carboxyl terminus and third intracellular loop. arrestin2 translocated significantly more rapidly than arrestin1 to agonist-activated membrane CRF₁ receptors in multiple cell lines. Although CRF₁ receptors internalized with agonist treatment, neither arrestin isoform trafficked with the receptor inside the cell, indicating that CRF₁ receptor-arrestin complexes dissociate at or near the cell membrane. Both arrestin- and clathrin-dependent mechanisms were involved in CRF₁ receptor internalization. To investigate molecular determinants mediating the robust arrestin2-CRF₁ receptor interaction, mutagenesis was performed to remove potential GRK phosphorylation sites. Truncating the CRF₁ receptor carboxyl terminus at serine-386 greatly reduced agonist-dependent phosphorylation, but only partially impaired arrestin2 recruitment. Removal of a serine/threonine cluster in the third intracellular loop also significantly reduced CRF₁ receptor phosphorylation, but did not alter arrestin2 recruitment. Phosphorylation was abolished in a CRF₁ receptor possessing both mutations. Surprisingly, this mutant still recruited arrestin2. These mutations did not alter membrane expression or cyclic AMP signaling of CRF₁ receptors. Our data reveal the involvement of at least two distinct receptor regions in arrestin2 recruitment: (i) a carboxyl-terminal motif in which serine/threonine residues must be phosphorylated; and (ii) an intracellular loop motif configured by agonist-induced changes in CRF₁ receptor conformation. Deficient arrestin2-CRF₁ receptor interactions could contribute to the pathophysiology of affective disorders by inducing excessive CRF₁ receptor signaling.