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Gene Synthesis | Multiple rounds of site-directed mutagenesis were performed to generate SSTR3-cK0, a variant with all the cytoplasm-facing lysine residues (K233, K330, K356, K407, and K421) mutated to arginine. GPR161-cK0, a variant with 18 cytoplasm-exposed lysine residues (K83, K84, K93, K167, K247, K250, K269, K296, K358, K362, K455, K469, K473, K481, K486, K497, K504, and K548) mutated to arginine, was gene synthesized (GenScript). | Get A Quote |
G-protein coupled receptors (GPCRs) undergo regulated trafficking into and out of cilia to control cilium-based signaling pathways. β-arrestin 2, a molecular sensor of activated GPCRs, and the BBSome are required for the signal-dependent exit of ciliary GPCRs but it is not known how β-arrestin 2 relays the activation state of GPCRs to the ciliary exit machinery. Here we find that, upon activation, the ciliary GPCRs SSTR3 and GPR161 become tagged with K63-linked ubiquitin (K63Ub) chains in a β-arrestin 2-dependent manner prior to BBSome-mediated exit. Removal of ubiquitin acceptor residues from SSTR3 and GPR161 demonstrates that ubiquitination of ciliary GPCRs is required for their regulated exit from cilia. ... More