TY - JOUR T1 - Lysine63-linked ubiquitin chains earmark GPCRs for BBSome-mediated removal from cilia JF - bioRxiv DO - 10.1101/2020.03.04.977090 SP - 2020.03.04.977090 AU - Swapnil Rohidas Shinde AU - Andrew R. Nager AU - Maxence V. Nachury Y1 - 2020/01/01 UR - http://biorxiv.org/content/early/2020/09/28/2020.03.04.977090.abstract N2 - Regulated trafficking of G-protein coupled receptors (GPCRs) controls cilium-based signaling pathways. β-arrestin, a molecular sensor of activated GPCRs, and the BBSome, a complex of Bardet-Biedl Syndrome (BBS) proteins, are required for the signal-dependent exit of ciliary GPCRs but the functional interplay between β-arrestin and the BBSome remains elusive. Here we find that, upon activation, ciliary GPCRs become tagged with K63-linked ubiquitin (K63Ub) chains in a β-arrestin-dependent manner prior to BBSome-mediated exit. Removal of ubiquitin acceptor residues from the somatostatin receptor 3 (SSTR3) and from the orphan GPCR GPR161 demonstrates that ubiquitination of ciliary GPCRs is required for their regulated exit from cilia. Furthermore, targeting a K63Ub-specific deubiquitinase to cilia blocks the exit of GPR161, SSTR3 and Smoothened (SMO) from cilia. Finally, ubiquitinated proteins accumulate in cilia of mammalian photoreceptors and Chlamydomonas cells when BBSome function is compromised. We conclude that K63Ub chains mark GPCRs and other unwanted ciliary proteins for recognition by the ciliary exit machinery.Competing Interest StatementThe authors have declared no competing interest. ER -