title: Both K63 and K48 ubiquitin linkages signal lysosomal degradation of the LDL receptor.
abstract: Linkage-specific ubiquitination often leads to distinct cellular events. It has been difficult to establish definitively the requirement for a particular linkage in mammalian degradation pathways due to the inability to deplete endogenous ubiquitin while maintaining cell viability. The E3 ubiquitin ligase inducible degrader of the LDL receptor (IDOL) targets the low density lipoprotein receptor (LDLR) for degradation. The nature of the linkages employed to signal lysosomal degradation of the LDLR, and to signal proteasomal autodegradation of IDOL, have not been determined. We used an inducible RNAi strategy to replace endogenous ubiquitin with mutants lacking K48 or K63. We found that IDOL catalyzes the transfer of ubiquitin chains to itself and to the LDLR that do not contain exclusively K48 or K63 linkages. Thus, LDLR can be targeted to the lysosome by either K48 or K63 linkages. We further demonstrate that although both ubiquitin conjugating enzyme E2 (UBE2)Ds and UBE2N/V1 can catalyze LDLR ubiquitination in a cell-free system, UBE2Ds appear to be the major E2 enzymes employed by IDOL in cells, consistent with their ability to catalyze both K48 and K63 linkages. The results reveal mechanistic insight into the posttranscriptional control of lipoprotein uptake and provide a test of the requirement of linkage-specific ubiquitination for specific lysosomal and proteasomal degradation pathways in mammalian cells.
authors: Howard Hughes Medical Institute and Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California at Los Angeles, Los Angeles, CA 90095, USA.
authors: Li
authors: L
authors: Zhang
authors: Ming
authors: M
authors: Xu
authors: Elena
authors: E
authors: Scotti
authors: Zhijian J
authors: ZJ
authors: Chen
authors: Peter
authors: P
authors: Tontonoz
mesh_terms: D002474:Cell-Free System; D004705:Endocytosis; D057809:HEK293 Cells; D006801:Humans; D008074:Lipoproteins / Q000378:metabolism; D008247:Lysosomes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D046988:Proteasome Endopeptidase Complex / Q000235:genetics / Q000378:metabolism; D059748:Proteolysis; D011973:Receptors, LDL / Q000737:chemistry / Q000378:metabolism*; D044767:Ubiquitin-Protein Ligases / Q000737:chemistry / Q000378:metabolism*; D054875:Ubiquitination / Q000235:genetics
chemical_list: C573412:LDLR protein, human; D008074:Lipoproteins; D011973:Receptors, LDL; D044767:Ubiquitin-Protein Ligases; D046988:Proteasome Endopeptidase Complex