SCAMP3 Negatively Regulates Epidermal Growth Factor Receptor Degradation and Promotes Receptor Recycling

被引:53
作者
Aoh, Quyen L. [1 ]
Castle, Anna M. [1 ]
Hubbard, Charles H. [1 ]
Katsumata, Osamu [2 ]
Castle, J. David [1 ]
机构
[1] Univ Virginia, Dept Cell Biol, Charlottesville, VA 22908 USA
[2] Nihon Univ, Sch Dent Matsudo, Dept Physiol, Matsudo, Chiba 2718587, Japan
基金
美国国家卫生研究院;
关键词
CARRIER MEMBRANE-PROTEINS; MULTIVESICULAR BODY FORMATION; CORE VESICLE EXOCYTOSIS; EPITHELIAL NA+ CHANNEL; UBIQUITIN LIGASE; DOWN-REGULATION; EARLY ENDOSOMES; EGF RECEPTOR; INWARD VESICULATION; ESCRT MACHINERY;
D O I
10.1091/mbc.E08-09-0894
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
The epidermal growth factor receptor (EGFR) is targeted for lysosomal degradation by ubiquitin-mediated interactions with the ESCRTs (endosomal-sorting complexes required for transport) in multivesicular bodies (MVBs). We show that secretory carrier membrane protein, SCAMP3, localizes in part to early endosomes and negatively regulates EGFR degradation through processes that involve its ubiquitylation and interactions with ESCRTs. SCAMP3 is multimonoubiquitylated and is able to associate with Nedd4 HECT ubiquitin ligases and the ESCRT-I subunit Tsg101 via its PY and PSAP motifs, respectively. SCAMP3 also associates with the ESCRT-0 subunit Hrs. Depletion of SCAMP3 in HeLa cells by inhibitory RNA accelerated degradation of EGFR and EGF while inhibiting recycling. Conversely, overexpression enhanced EGFR recycling unless ubiquitylatable lysines, PY or PSAP motifs in SCAMP3 were mutated. Notably, dual depletions of SCAMP3 and ESCRT subunits suggest that SCAMP3 has a distinct function in parallel with the ESCRTs that regulates receptor degradation. This function may affect trafficking of receptors from prelysosomal compartments as SCAMP3 depletion appeared to sustain the incidence of EGFR-containing MVBs detected by immunoelectron microscopy. Together, our results suggest that SCAMP3, its modification with ubiquitin, and its interactions with ESCRTs coordinately regulate endosomal pathways and affect the efficiency of receptor down-regulation.
引用
收藏
页码:1816 / 1832
页数:17
相关论文
共 90 条
[1]   UBPY-mediated epidermal growth factor receptor (EGFR) de-ubiquitination promotes EGFR degradation [J].
Alwan, Husam A. J. ;
van Leeuwen, Jeroen E. M. .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2007, 282 (03) :1658-1669
[2]   Tal, a Tsg101-specific E3 ubiquitin ligase, regulates receptor endocytosis and retrovirus budding [J].
Amit, I ;
Yakir, L ;
Katz, M ;
Zwang, Y ;
Marmor, MD ;
Citri, A ;
Shtiegman, K ;
Alroy, I ;
Tuvia, S ;
Reiss, Y ;
Roubini, E ;
Cohen, M ;
Wides, R ;
Bacharach, E ;
Schubert, U ;
Yarden, Y .
GENES & DEVELOPMENT, 2004, 18 (14) :1737-1752
[3]   A protein's final ESCRT [J].
Babst, M .
TRAFFIC, 2005, 6 (01) :2-9
[4]   Hrs regulates multivesicular body formation via ESCRT recruitment to endosomes [J].
Bache, KG ;
Brech, A ;
Mehlum, A ;
Stenmark, H .
JOURNAL OF CELL BIOLOGY, 2003, 162 (03) :435-442
[5]   STAM and Hrs are subunits of a multivalent ubiquitin-binding complex on early endosomes [J].
Bache, KG ;
Raiborg, C ;
Mehlum, A ;
Stenmark, H .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2003, 278 (14) :12513-12521
[6]   The ESCRT-III subunit hVps24 is required for degradation but not silencing of the epidermal growth factor receptor [J].
Bache, Kristi G. ;
Stuffers, Susanne ;
Malerod, Lene ;
Slagsvold, Thomas ;
Raiborg, Camilla ;
Lechardeur, Delphine ;
Walchli, Sebastien ;
Lukacs, Gergely L. ;
Brech, Andreas ;
Stenmark, Harald .
MOLECULAR BIOLOGY OF THE CELL, 2006, 17 (06) :2513-2523
[7]   Plasticity of polyubiquitin recognition as lysosomal targeting signals by the endosomal sorting machinery [J].
Barriere, Herve ;
Nemes, Csilla ;
Du, Kai ;
Lukacs, Gergely L. .
MOLECULAR BIOLOGY OF THE CELL, 2007, 18 (10) :3952-3965
[8]   Degradation of endocytosed epidermal growth factor and virally ubiquitinated major histocompatibility complex class I is independent of mammalian ESCRTII [J].
Bowers, K ;
Piper, SC ;
Edeling, MA ;
Gray, SR ;
Owen, DJ ;
Lehner, PJ ;
Luzio, JP .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2006, 281 (08) :5094-5105
[9]  
BRAND SH, 1991, J BIOL CHEM, V266, P18949
[10]   Regulation of epidermal growth factor receptor signaling by endocytosis and intracellular trafficking [J].
Burke, P ;
Schooler, K ;
Wiley, HS .
MOLECULAR BIOLOGY OF THE CELL, 2001, 12 (06) :1897-1910