title: The role of ubiquitination in lysosomal trafficking of δ-opioid receptors.
abstract: The δ-opioid receptor (DOR) undergoes ligand-induced downregulation by endosomal sorting complex required for transport (ESCRT)-dependent endocytic trafficking to lysosomes. In contrast to a number of other signaling receptors, the DOR can downregulate effectively when its ubiquitination is prevented. We explored the membrane trafficking basis of this behavior. First, we show that internalized DORs traverse the canonical multivesicular body (MVB) pathway and localize to intralumenal vesicles (ILVs). Second, we show that DOR ubiquitination stimulates, but is not essential for, receptor transfer to ILVs and proteolysis of the receptor endodomain. Third, we show that receptor ubiquitination plays no detectable role in the early sorting of internalized DORs out of the recycling pathway. Finally, we show that DORs undergo extensive proteolytic fragmentation in the ectodomain, even when receptor ubiquitination is prevented or ILV formation itself is blocked. Together, these results are sufficient to explain why DORs downregulate effectively in the absence of ubiquitination, and they place a discrete molecular sorting operation in the MVB pathway effectively upstream of the ESCRT. More generally, these findings support the hypothesis that mammalian cells can control the cytoplasmic accessibility of internalized signaling receptors independently from their ultimate trafficking fate.
authors.affiliation: Program in Cell Biology, Department of Biochemistry and Biophysics, University of California, San Francisco, CA, USA.
authors.forename: Anastasia G
authors.initials: AG
authors.lastname: Henry
authors.forename: Ian J
authors.initials: IJ
authors.lastname: White
authors.forename: Mark
authors.initials: M
authors.lastname: Marsh
authors.forename: Mark
authors.initials: M
authors.lastname: von Zastrow
authors.forename: James N
authors.initials: JN
authors.lastname: Hislop
mesh_terms: D002461:Cell Line, Transformed; D002478:Cells, Cultured; D015536:Down-Regulation; D004705:Endocytosis / Q000235:genetics / Q000502:physiology; D056827:Endosomal Sorting Complexes Required for Transport / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D008247:Lysosomes / Q000235:genetics / Q000378:metabolism*; D057146:Multivesicular Bodies / Q000235:genetics / Q000378:metabolism*; D021381:Protein Transport; D017465:Receptors, Opioid, delta / Q000235:genetics / Q000378:metabolism*; D025801:Ubiquitin / Q000378:metabolism*; D054875:Ubiquitination / Q000502:physiology*
chemical_list: D056827:Endosomal Sorting Complexes Required for Transport; D017465:Receptors, Opioid, delta; D025801:Ubiquitin