Involvement of the ubiquitin/proteasome system in sorting of the interleukin 2 receptor beta chain to late endocytic compartments.
Down-regulation of cell surface growth factor receptors plays a key role in the tight control of cellular responses. Recent reports suggest that the ubiquitin system, in addition to participating in degradation by the proteasome of cytosolic and nuclear proteins, might also be involved in the down-regulation of various membrane receptors. We have previously characterized a signal in the cytosolic part of the interleukin 2 receptor beta chain (IL2Rbeta) responsible for its targeting to late endosomes/lysosomes. In this report, the role of the ubiquitin/proteasome system on the intracellular fate of IL2Rbeta was investigated. Inactivation of the cellular ubiquitination machinery in ts20 cells, which express a thermolabile ubiquitin-activating enzyme E1, leads to a significant decrease in the degradation rate of IL2Rbeta, with little effect on its internalization. In addition, we show that a fraction of IL2Rbeta can be monoubiquitinated. Furthermore, mutation of the lysine residues of the cytosolic region of a chimeric receptor carrying the IL2Rbeta targeting signal resulted in a decreased degradation rate. When cells expressing IL2Rbeta were treated either by proteasome or lysosome inhibitors, a significant decrease in receptor degradation was observed. Our data show that ubiquitination is required for the sorting of IL2Rbeta toward degradation. They also indicate that impairment of proteasome function might more generally affect intracellular routing.
Rocca
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Unité de Biologie des Interactions Cellulaires, Unité de Recherche Associée Centre National de la Recherche Scientifique 1960, Institut Pasteur, 75724 Paris Cedex 15, France.
Lamaze
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Subtil
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Dautry-Varsat
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D000111:Acetylcysteine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000962:Antimalarials / Q000494:pharmacology; D002460:Cell Line; D002738:Chloroquine / Q000494:pharmacology; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004705:Endocytosis / Q000502:physiology*; D006801:Humans; D015151:Immunoblotting; D007976:Leupeptins / Q000494:pharmacology; D008856:Microscopy, Fluorescence; D009097:Multienzyme Complexes / Q000037:antagonists & inhibitors / Q000378:metabolism*; D046988:Proteasome Endopeptidase Complex; D021382:Protein Sorting Signals / Q000502:physiology*; D021122:Protein Subunits; D015375:Receptors, Interleukin-2 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D014452:Ubiquitins / Q000378:metabolism*
D000962:Antimalarials; D015853:Cysteine Proteinase Inhibitors; D007976:Leupeptins; D009097:Multienzyme Complexes; D021382:Protein Sorting Signals; D021122:Protein Subunits; D015375:Receptors, Interleukin-2; D011993:Recombinant Fusion Proteins; D014452:Ubiquitins; C067713:lactacystin; D002738:Chloroquine; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex; C032854:leupeptin; D000111:Acetylcysteine