Regulation of Epidermal Growth Factor Receptor Down-Regulation by UBPY-mediated Deubiquitination at Endosomes

Mizuno, Emi; Iura, Takanobu; Mukai, Akiko; Yoshimori, Tamotsu; Kitamura, Naomi; Komada, Masayuki

doi:10.1091/mbc.e05-06-0560

Cited by 249 publications

(288 citation statements)

References 55 publications

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“…In agreement with this role, we observe that AMSH D348A induces an increased ubiquitination of an endosomal cargo, namely MLV Gag. A role of UBPY in the deubiquitination of endosomal cargo in transit to the MVB has also been proposed (48), but there are important differences with AMSH. The activity of UBPY seems to be dependent of the interaction with STAM/HRS (48), whereas the dominant-negative activity of AMSH and its interaction with ESCRT-III are independent of STAM1 binding.…”

Section: Discussionmentioning

confidence: 99%

“…The activity of UBPY seems to be dependent of the interaction with STAM/HRS (48), whereas the dominant-negative activity of AMSH and its interaction with ESCRT-III are independent of STAM1 binding. Interestingly, AMSH has been shown to process K63-linked polyubiquitin (26,48), which regulates endocytosis and lysosomal sorting (49) whereas UBPY processes K48-linked polyubiquitin, a signal for proteasomal degradation.…”

Section: Discussionmentioning

confidence: 99%

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Interaction of AMSH with ESCRT-III and Deubiquitination of Endosomal Cargo

Agromayor

Martín-Serrano

2006

Journal of Biological Chemistry

116

117

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The "class E" vacuolar protein sorting (VPS) pathway mediates sorting of ubiquitinated cargo into the forming vesicles of the multivesicular bodies (MVB), and it is essential for down-regulation of signaling by growth factors and budding of enveloped viruses such as Ebola and HIV-1. Work in yeast has identified DOA4 as a gene that is recruited by the class E machinery to remove ubiquitin from the endosomal cargo before it is incorporated into MVB vesicles, but the identity of the mammalian counterpart is unclear. Here we report the interaction of AMSH (associated molecule with the SH3 domain of STAM), an endosomal deubiquitinating enzyme, with the endodomal sorting complex required for transport (ESCRT-III) subunits CHMP1A, CHMP1B, CHMP2A, and CHMP3. We also show that a catalytically inactive AMSH inhibits retroviral budding in a dominant-negative manner and induces the accumulation of ubiquitinated forms of an endosomal cargo, namely murine leukemia virus Gag. Finally, VPS4 and AMSH compete for binding to the C-terminal regions of CHMP1A and CHMP1B, revealing a coordinated interaction with ESCRT-III. Taken together, these results are consistent with a role of AMSH in the deubiquitination of the endosomal cargo preceding lysosomal degradation.Ubiquitin plays an essential role in the trafficking of membrane proteins into the degradative lysosomal pathway. Covalent attachment of ubiquitin to certain activated cell surface receptors serves as a sorting signal for entry into the endocytic vesicles at the plasma membrane (1). Subsequently to this internalization, the class E vacuolar protein sorting (VPS) 2 pathway directs the ubiquitinated cargo into the forming vesicles of the late endosomal compartment, termed multivesicular bodies (MVB) (2, 3). In the last step, the MVB fuses with the lysosomal membrane and the cargo is delivered to the lumen of the lysosome for degradation.Work in yeast has identified 18 class E VPS proteins that are required for sorting into the lumen of MVBs (2, 4, 5). Importantly, at least one human homolog for every class E protein has been identified and recent studies show that most of the protein-protein interactions between the class E proteins are conserved from yeasts to humans (6 -8). A subset of class E proteins form one of three endosomal sorting complexes required for transport (ESCRT-I, -II, and -III) (9 -11), and current models propose that these complexes are sequentially recruited by the endosomal cargo, forming a membrane-associated lattice that functions in vesicle invagination during MVB biogenesis. ESCRT-III is believed to contain the core sorting machinery of the class E pathway and it is composed of two functional subcomplexes, a membrane-proximal complex that interacts with the endosomal membrane and a peripheral subcomplex that seems to recruit accessory proteins (9).After completion of cargo sorting and vesicle formation, ESCRT-III recruits the AAA ATPase Vps4 through direct protein-protein interactions, mediating the disassembly of the ESCRT machinery for recycling in...

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Interaction of AMSH with ESCRT-III and Deubiquitination of Endosomal Cargo

Agromayor

Martín-Serrano

2006

Journal of Biological Chemistry

116

117

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“…One possible explanation is that there may be a redundant DUB activity in mammals. Recently, a DUB called UBPY was suspected as a possible Doa4 orthologue in mammals, but the same report showed that the amount of free ubiquitin in UBPYdepleted cells was unaltered (Mizuno et al, 2005). In addition, there is no evidence so far supporting UBPY's involvement in ESCRT-III.…”

Section: Discussionmentioning

confidence: 99%

“…In vitro deubiquitination assay K48-linked or K63-linked polyubiquitin chains (500 ng; Boston Biochem) were incubated with 5 µg of GST-fusion proteins at 37°C for 18 hours in DUB buffer (PBS [pH 7.0], 5 mM MgCl2, 2 mM DTT) (Mizuno et al, 2005). Tris-Tricine Sample Buffer (0.1 M Tris-HCl [pH 6.8], 24% Glycerol, 1% SDS, 2% β-mercaptoethanol, and 2% Coomassie G-250) was added, and the reaction mixture was heated to 95°C for 5 minutes and resolved on 16.5% Ready Gels J Peptide (BioRad).…”

Section: Preparation Of Cytosol and Membrane Fractionsmentioning

confidence: 99%

AMSH, an ESCRT-III Associated Enzyme, Deubiquitinates Cargo on MVB/Late Endosomes

Kyuuma

Kikuchi

Kojima

et al. 2006

Cell Struct. Funct.

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and 6 These two authors contributed equally to this work.ABSTRACT. The appropriate sorting of vesicular cargo, including cell-surface proteins, is critical for many cellular functions. Ubiquitinated cargo is targeted to endosomes and digested by lysosomal enzymes. We previously identified AMSH, a deubiquitination enzyme (DUB), to be involved in vesicular transport. Here, we purified an AMSH-binding protein, CHMP3, which is an ESCRT-III subunit. ESCRT-III functions on maturing endosomes, indicating AMSH might also play a role in MVB/late endosomes. Expression of an AMSH mutant lacking CHMP3-binding ability resulted in aberrant endosomes with accumulations of ubiquitinated cargo. Nevertheless, CHMP3-binding capability was not essential for AMSH's in vitro DUB activity or its endosomal localization, suggesting that, in vivo, the deubiquitination of endosomal cargo is CHMP3-dependent. Ubiquitinated cargo also accumulated on endosomes when catalytically inactive AMSH was expressed or AMSH was depleted. These results suggest that both the DUB activity of AMSH and its CHMP3-binding ability are required to clear ubiquitinated cargo from endosomes.

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“…The best candidate mammalian Doa4 ortholog is Usp8/UBPY, which can also bind ESCRT-III; however, its role in rescuing Ub from lysosomal degradation has not been specifically determined (Wright et al 2011). Indeed, USP8/ UBPY as well as a similarly positioned DUb, AMSH, can bind both ESCRT-0 and ESCRT-III and a host of experiments looking at different cargoes suggest they can play a role in deubiquitinating cargo before MVB sorting to promote cargo recycling (McCullough et al 2004;Mizuno et al 2005;Bowers et al 2006;Berlin et al 2010;Zhou et al 2013), cargo deubiquitination after sorting as a way to release them from the ESCRT apparatus and promote sorting into MVBs (Alwan and van Leeuwen 2007;Balut et al 2011), or as a way of deubiquitinating ESCRT components to rescue them from degradation or inactive conformations (Row et al 2007;Sierra et al 2010). One of the interesting aspects of cargo deubiquitination in the final stages of sorting is that it predicts that cargo is sorted into a domain capable of trapping Ub cargo such that it cannot escape upon deubiquitination.…”

Section: Ubiquitin-dependent Sorting In Endocytosismentioning

confidence: 99%

Ubiquitin-Dependent Sorting in Endocytosis

Piper

Đikić

Lukacs

2014

Cold Spring Harbor Perspectives in Biology

182

171

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When ubiquitin (Ub) is attached to membrane proteins on the plasma membrane, it directs them through a series of sorting steps that culminate in their delivery to the lumen of the lysosome where they undergo complete proteolysis. Ubiquitin is recognized by a series of complexes that operate at a number of vesicle transport steps. Ubiquitin serves as a sorting signal for internalization at the plasma membrane and is the major signal for incorporation into intraluminal vesicles of multivesicular late endosomes. The sorting machineries that catalyze these steps can bind Ub via a variety of Ub-binding domains. At the same time, many of these complexes are themselves ubiquitinated, thus providing a plethora of potential mechanisms to regulate their activity. Here we provide an overview of how membrane proteins are selected for ubiquitination and deubiquitination within the endocytic pathway and how that ubiquitin signal is interpreted by endocytic sorting machineries. HOW PROTEINS ARE SELECTED FOR UBIQUITINATIONU biquitin (Ub) is covalently attached to substrate proteins by the concerted action of E2-conjugating enzymes and E3 ligases (Varshavsky 2012). Most of the specificity and regulation of ubiquitination is at the level of the E3 ligases, which vastly outnumber the E2-conjugating enzymes. Ubiquitination results from the transfer of Ub, held either by the E2 or in some cases by the E3 as a high-energy thioester bond, onto a substrate protein, typically on the terminal amide of a substrate acceptor lysine side chain. Owing to the intense interest in the ubiquitination system, many of the simple rules and distinctions concerning different types of ligases, their specificity for forming particular polyUb chains, and even the kinds of residues in substrate proteins that can be ubiquitin modified, have been blurred with the discovery of mixed poly-Ub chains, new enzymatic mechanisms for Ub transfer, and ubiquitination of nonlysine residues (Kulathu and Komander 2012;Wenzel and Klevit 2012;McDowell and Philpott 2013). Nonetheless, in basic terms, Ub ligases function as platforms that coordinate substrate recognition with Ub transfer as well as configuring poly-Ub chain topology by the E2-conjugating enzyme. Substrates can be bound directly by the E3 ligase or by adaptor proteins that in turn bind to ligases, and selecEditors:

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Regulation of Epidermal Growth Factor Receptor Down-Regulation by UBPY-mediated Deubiquitination at Endosomes

Cited by 249 publications

References 55 publications

Interaction of AMSH with ESCRT-III and Deubiquitination of Endosomal Cargo

Interaction of AMSH with ESCRT-III and Deubiquitination of Endosomal Cargo

AMSH, an ESCRT-III Associated Enzyme, Deubiquitinates Cargo on MVB/Late Endosomes

Ubiquitin-Dependent Sorting in Endocytosis

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