A single ubiquitin is sufficient for cargo protein entry into MVBs in the absence of ESCRT ubiquitination

Stringer, Daniel K.; Piper, Robert C.

doi:10.1083/jcb.201008121

Cited by 140 publications

(180 citation statements)

References 72 publications

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“…To this end, we expressed a dominantacting protein in which the ESCRT-0 subunit Hse1 (the homologue of mammalian STAM) is fused to the catalytic domain of the deubiquitinating peptidase domain of Herpes Virus UL36, resulting in the formation of an Hse1-DUb chimeric protein ( Fig 1A). Our previous studies showed that when Hse1-DUb is expressed in wild-type cells, either as the sole copy of Hse1 or in addition to endogenous Hse1, it effectively removes Ub from cargo during the MVB-sorting process, and blocks the sorting of a variety of cargoes that would normally undergo Ub-dependent sorting into MVB ILVs [7]. As a further measure of the effectiveness of Hse1-DUb to block MVB sorting of ubiquitinated cargo, we followed green fluorescent protein (GFP)-tagged Ub (GFP-Ub; [8]).…”

Section: Resultsmentioning

confidence: 99%

Cargo ubiquitination is essential for multivesicular body intralumenal vesicle formation

et al. 2012

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The efficient formation of a variety of transport vesicles is influenced by the presence of cargo, suggesting that cargo itself might have a defining role in vesicle biogenesis. However, definitive in vivo experiments supporting this concept are lacking, as it is difficult to eliminate endogenous cargo. The Endosomal Sorting Complexes Required for Transport (ESCRT) apparatus sorts ubiquitinated membrane proteins into endosomal intralumenal vesicles (ILVs) that accumulate within multivesicular bodies. Here we show that cargo ubiquitination is required for effective recruitment of the ESCRT machinery onto endosomal membranes and for the subsequent formation of ILVs.

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Section: Resultsmentioning

confidence: 99%

Cargo ubiquitination is essential for multivesicular body intralumenal vesicle formation

et al. 2012

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“…Ubiquitylation of PM proteins has been demonstrated in higher plants (6)(7)(8)(9)15), and, analogous to the situation in animals and fungi (30)(31)(32), covalent attachment of a single ubiquitin seems a sufficient signal for endocytosis and vacuolar targeting of selected PM proteins (7,8). This modification has been implicated in modulating abundance of nutrient carrier and light receptor proteins at the PM, thereby controlling intracellular solute homeostasis as well as stimulus-mediated growth responses (6)(7)(8).…”

Section: Resultsmentioning

confidence: 99%

Lysine ⁶³ -linked ubiquitylation of PIN2 auxin carrier protein governs hormonally controlled adaptation of Arabidopsis root growth

Leitner

Petrášek²,

Tomanov³

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

187

220

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Cross-talk between plant cells and their surroundings requires tight regulation of information exchange at the plasma membrane (PM), which involves dynamic adjustments of PM protein localization and turnover to modulate signal perception and solute transport at the interface between cells and their surroundings. In animals and fungi, turnover of PM proteins is controlled by reversible ubiquitylation, which signals endocytosis and delivery to the cell's lytic compartment, and there is emerging evidence for related mechanisms in plants. Here, we describe the fate of Arabidopsis PIN2 protein, required for directional cellular efflux of the phytohormone auxin, and identify cis-and trans-acting mediators of PIN2 ubiquitylation. We demonstrate that ubiquitin acts as a principal signal for PM protein endocytosis in plants and reveal dynamic adjustments in PIN2 ubiquitylation coinciding with variations in vacuolar targeting and proteolytic turnover. We show that control of PIN2 proteolytic turnover via its ubiquitylation status is of significant importance for auxin distribution in root meristems and for environmentally controlled adaptations of root growth. Moreover, we provide experimental evidence indicating that PIN2 vacuolar sorting depends on modification specifically by lysine 63 -linked ubiquitin chains. Collectively, our results establish lysine 63 -linked PM cargo ubiquitylation as a regulator of polar auxin transport and adaptive growth responses in higher plants.P lants have evolved a repertoire of mechanisms for continuously adapting vital parameters in response to fluctuating environmental conditions. Sensing and responding to such variations depend to a large extent on the activity of plasma membrane (PM)-associated proteins that function in stimulus perception and solute transport. Specifically, adjustments in subcellular distribution of PM proteins are an efficient means to modulate their activity and involve continuous protein cycling between PM and endosomes as well as irreversible targeting for degradation in the lytic vacuole/lysosome (1).An evolutionary conserved machinery controls PM protein sorting for degradation, and, specifically in animals and fungi, it was demonstrated that PM protein fate is decisively influenced by their reversible ubiquitylation, triggering cargo endocytosis and delivery to the lytic compartment (2-4). Related mechanisms appear to be operative in plants (5-9) because, recently, ubiquitylation has been described for some plant PM proteins, linking nutrient transport and stimulus perception to endocytic protein turnover (6-9). Strikingly, different patterns of protein ubiquitylation have been observed, with mono-, di-, as well as polyubiquitylation implicated in regulating endocytic trafficking and degradation of distinct proteins (5). This resembles the situation in nonplant organisms, in which mono-and polyubiquitylation have been associated with cargo endocytosis (10).Vacuolar sorting was also demonstrated for PIN1-type auxin carrier proteins, which are instrumental for ...

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“…Specificity for K63-linked chains, as opposed to multiple-mono-Ub, has been suggested by the cooperative binding K63 poly-Ub shows for the multiple UBDs within ESCRT-0 or a single UBD within yeast Vps36 (Kulathu et al 2009;Lange et al 2012b). However, these effects are moderate and a single Ub on cargo is sufficient to confer sorting into ILVs, indicating that the role of K63 chains may be to simply increase the number of mono-Ub sorting signals attached to cargo (Ren and Hurley 2010;Stringer and Piper 2011).…”

Section: Roles Of Multiple Ubdsmentioning

confidence: 99%

“…In experimental settings, namely, where Ub is permanently attached cargo, a single Ub is sufficient for delivery to lysosomes (Shih et al 2000;Raiborg et al 2002;Stringer and Piper 2011). However, as an internalization signal, a single Ub is exceptionally weak and Ub is likely operational only in chains (Barriere et al 2006;Hawryluk et al 2006;Bertelsen et al 2011).…”

Section: Ubiquitin-dependent Sorting In Endocytosismentioning

confidence: 99%

Ubiquitin-Dependent Sorting in Endocytosis

Piper

Đikić

Lukacs

2014

Cold Spring Harbor Perspectives in Biology

193

176

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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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A single ubiquitin is sufficient for cargo protein entry into MVBs in the absence of ESCRT ubiquitination

Abstract: While ESCRT-0 is ubiquitinated by the Rsp5 E3 ligase, loss of Rsp5 does not disrupt monoubiquitin-dependent sorting into multivesicular bodies.

Cited by 140 publications

References 72 publications

Cargo ubiquitination is essential for multivesicular body intralumenal vesicle formation

Cargo ubiquitination is essential for multivesicular body intralumenal vesicle formation

Lysine ⁶³ -linked ubiquitylation of PIN2 auxin carrier protein governs hormonally controlled adaptation of Arabidopsis root growth

Ubiquitin-Dependent Sorting in Endocytosis

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A single ubiquitin is sufficient for cargo protein entry into MVBs in the absence of ESCRT ubiquitination

Abstract: While ESCRT-0 is ubiquitinated by the Rsp5 E3 ligase, loss of Rsp5 does not disrupt monoubiquitin-dependent sorting into multivesicular bodies.

Cited by 140 publications

References 72 publications

Cargo ubiquitination is essential for multivesicular body intralumenal vesicle formation

Cargo ubiquitination is essential for multivesicular body intralumenal vesicle formation

Lysine 63 -linked ubiquitylation of PIN2 auxin carrier protein governs hormonally controlled adaptation of Arabidopsis root growth

Ubiquitin-Dependent Sorting in Endocytosis

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Lysine ⁶³ -linked ubiquitylation of PIN2 auxin carrier protein governs hormonally controlled adaptation of Arabidopsis root growth