Vps22/EAP30 in ESCRT-II mediates endosomal sorting of growth factor and chemokine receptors destined for lysosomal degradation.
The ubiquitin-binding protein Hrs and endosomal sorting complex required for transport (ESCRT)-I and ESCRT-III are involved in sorting endocytosed and ubiquitinated receptors to lysosomes for degradation and efficient termination of signaling. In this study, we have investigated the role of the ESCRT-II subunit Vps22/EAP30 in degradative protein sorting of ubiquitinated receptors. Vps22 transiently expressed in HeLa cells was detected in endosomes containing endocytosed epidermal growth factor receptors (EGFRs) as well as Hrs and ESCRT-I and ESCRT-III. Depletion of Vps22 by small interfering RNA, which was accompanied by decreased levels of other ESCRT-II subunits, greatly reduced degradation of EGFR and its ligand EGF as well as the chemokine receptor CXCR4. EGFR accumulated on the limiting membranes of early endosomes and aberrantly small multivesicular bodies in Vps22-depleted cells. Phosphorylation and nuclear translocation of extracellular-signal-regulated kinase1/2 downstream of the EGF-activated receptor were sustained by depletion of Hrs or the ESCRT-I subunit Tsg101. In contrast, this was not the case when Vps22 was depleted. These results indicate an important role for Vps22 in ligand-induced EGFR and CXCR4 turnover and suggest that termination of EGF signaling occurs prior to ESCRT-II engagement.
Malerød
Lene
L
Centre for Cancer Biomedicine, University of Oslo, Montebello, N-0310 Oslo, Norway.
Stuffers
Susanne
S
Brech
Andreas
A
Stenmark
Harald
H
D056827:Endosomal Sorting Complexes Required for Transport; D011992:Endosomes / Q000378:metabolism*; D004815:Epidermal Growth Factor / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008247:Lysosomes / Q000378:metabolism*; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D010766:Phosphorylation; D010957:Plasmids / Q000378:metabolism; D021381:Protein Transport; D011506:Proteins / Q000737:chemistry / Q000502:physiology*; D034741:RNA, Small Interfering / Q000378:metabolism; D019707:Receptors, Chemokine / Q000378:metabolism*; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000502:physiology*; D015398:Signal Transduction; D025801:Ubiquitin / Q000378:metabolism
D056827:Endosomal Sorting Complexes Required for Transport; D011506:Proteins; D034741:RNA, Small Interfering; D019707:Receptors, Chemokine; D043562:Receptors, G-Protein-Coupled; C094045:SNF8 protein, S cerevisiae; C525859:SNF8 protein, human; D029701:Saccharomyces cerevisiae Proteins; D025801:Ubiquitin; D004815:Epidermal Growth Factor