scholarly journals Coordinated binding of Vps4 to ESCRT-III drives membrane neck constriction during MVB vesicle formation

2014 ◽  
Vol 205 (1) ◽  
pp. 33-49 ◽  
Author(s):  
Manuel Alonso Y Adell ◽  
Georg F. Vogel ◽  
Mehrshad Pakdel ◽  
Martin Müller ◽  
Herbert Lindner ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Adenosine Triphosphatase ◽  
Electron Tomography ◽  
Vesicle Formation ◽  
Multivesicular Bodies ◽  
Membrane Remodeling ◽  
Yeast Genetics ◽  
Endosomal Sorting ◽  
Distinct Membrane

Five endosomal sorting complexes required for transport (ESCRTs) mediate the degradation of ubiquitinated membrane proteins via multivesicular bodies (MVBs) in lysosomes. ESCRT-0, -I, and –II interact with cargo on endosomes. ESCRT-II also initiates the assembly of a ringlike ESCRT-III filament consisting of Vps20, Snf7, Vps24, and Vps2. The AAA–adenosine triphosphatase Vps4 disassembles and recycles the ESCRT-III complex, thereby terminating the ESCRT pathway. A mechanistic role for Vps4 in intraluminal vesicle (ILV) formation has been unclear. By combining yeast genetics, biochemistry, and electron tomography, we find that ESCRT-III assembly on endosomes is required to induce or stabilize the necks of growing MVB ILVs. Yet, ESCRT-III alone is not sufficient to complete ILV biogenesis. Rather, binding of Vps4 to ESCRT-III, coordinated by interactions with Vps2 and Snf7, is coupled to membrane neck constriction during ILV formation. Thus, Vps4 not only recycles ESCRT-III subunits but also cooperates with ESCRT-III to drive distinct membrane-remodeling steps, which lead to efficient membrane scission at the end of ILV biogenesis in vivo.

scholarly journals Bro1 stimulates Vps4 to promote intralumenal vesicle formation during multivesicular body biogenesis

2021 ◽  
Vol 220 (8) ◽  
Author(s):  
Chun-Che Tseng ◽  
Shirley Dean ◽  
Brian A. Davies ◽  
Ishara F. Azmi ◽  
Natalya Pashkova ◽  
...  
Keyword(s):  
Multivesicular Body ◽  
Vesicle Formation ◽  
Multivesicular Bodies ◽  
Membrane Remodeling ◽  
Endosomal Sorting ◽  
Aaa Atpase ◽  
Ubiquitin Binding ◽  
Stimulation Of ◽  
Cargo Sorting

Endosomal sorting complexes required for transport (ESCRT-0, -I, -II, -III) execute cargo sorting and intralumenal vesicle (ILV) formation during conversion of endosomes to multivesicular bodies (MVBs). The AAA-ATPase Vps4 regulates the ESCRT-III polymer to facilitate membrane remodeling and ILV scission during MVB biogenesis. Here, we show that the conserved V domain of ESCRT-associated protein Bro1 (the yeast homologue of mammalian proteins ALIX and HD-PTP) directly stimulates Vps4. This activity is required for MVB cargo sorting. Furthermore, the Bro1 V domain alone supports Vps4/ESCRT–driven ILV formation in vivo without efficient MVB cargo sorting. These results reveal a novel activity of the V domains of Bro1 homologues in licensing ESCRT-III–dependent ILV formation and suggest a role in coordinating cargo sorting with membrane remodeling during MVB sorting. Moreover, ubiquitin binding enhances V domain stimulation of Vps4 to promote ILV formation via the Bro1–Vps4–ESCRT-III axis, uncovering a novel role for ubiquitin during MVB biogenesis in addition to facilitating cargo recognition.

scholarly journals Bro1 directly stimulates Vps4 activity to promote Intralumenal Vesicle Formation during Multivesicular Body biogenesis

2020 ◽  
Author(s):  
Chun-che Tseng ◽  
Shirley Dean ◽  
Brian A. Davies ◽  
Ishara F. Azmi ◽  
Natalya Pashkova ◽  
...  
Keyword(s):  
Multivesicular Body ◽  
Vesicle Formation ◽  
Multivesicular Bodies ◽  
Membrane Remodeling ◽  
Endosomal Sorting ◽  
Aaa Atpase ◽  
Evolutionarily Conserved ◽  
Carboxyl Terminal ◽  
Yeast Homolog ◽  
Direct Regulation

AbstractEndosomal sorting complexes required for transport (ESCRT-0, -I, -II, -III) form intralumenal vesicles (ILVs) during the conversion of endosomes to multivesicular bodies (MVBs). The assembly and disassembly of an ESCRT-III polymer facilitates membrane remodeling and scission during this process. The ESCRT-III-associated protein Bro1 (the yeast homolog of mammalian proteins ALIX and HD-PTP) promotes ESCRT-III assembly and inhibits disassembly of ESCRT-III filaments by impeding Vps4, a AAA-ATPase that dismantles ESCRT-III polymers. Here we show that the evolutionarily conserved “V domain” of Bro1-family proteins directly stimulate Vps4 ATPase activity and this activity is enhanced by interaction with ubiquitin. Surprisingly, a carboxyl-terminal fragment of Bro1 containing the V domain supports ILV formation but not sorting of cargo into ILVs, revealing that these two processes can be uncoupled. These studies implicate Bro1 as a factor coordinating cargo sorting with direct regulation of Vps4 to modulate ESCRT-III driven ILV formation during MVB biogenesis.

scholarly journals Fission of SNX-BAR–coated endosomal retrograde transport carriers is promoted by the dynamin-related protein Vps1

2014 ◽  
Vol 204 (5) ◽  
pp. 793-806 ◽  
Author(s):  
Richard J. Chi ◽  
Jingxuan Liu ◽  
Matthew West ◽  
Jing Wang ◽  
Greg Odorizzi ◽  
...  
Keyword(s):  
Retrograde Transport ◽  
Related Protein ◽  
Membrane Remodeling ◽  
Functional Link ◽  
Bar Domain ◽  
Endosomal Sorting ◽  
Sorting Nexin ◽  
Bar Domain Proteins

Retromer is an endosomal sorting device that orchestrates capture and packaging of cargo into transport carriers coated with sorting nexin BAR domain proteins (SNX-BARs). We report that fission of retromer SNX-BAR–coated tubules from yeast endosomes is promoted by Vps1, a dynamin-related protein that localizes to endosomes decorated by retromer SNX-BARs and Mvp1, a SNX-BAR that is homologous to human SNX8. Mvp1 exhibits potent membrane remodeling activity in vitro, and it promotes association of Vps1 with the endosome in vivo. Retrograde transport carriers bud from the endosome coated by retromer and Mvp1, and cargo export is deficient in mvp1- and vps1-null cells, but with distinct endpoints; cargo export is delayed in mvp1-null cells, but cargo export completely fails in vps1-null cells. The results indicate that Mvp1 promotes Vps1-mediated fission of retromer- and Mvp1-coated tubules that bud from the endosome, revealing a functional link between the endosomal sorting and fission machineries to produce retrograde transport carriers.

scholarly journals Revisiting caveolin trafficking: the end of the caveosome

2010 ◽  
Vol 191 (3) ◽  
pp. 439-441 ◽  
Author(s):  
Robert G. Parton ◽  
Mark T. Howes
Keyword(s):  
Membrane Proteins ◽  
Multivesicular Bodies ◽  
Endosomal Sorting ◽  
Cell Biol ◽  
New Findings

In this issue, a study by Hayer et al. (2010. J. Cell Biol. doi: 10.1083/jcb.201003086) provides insights into the trafficking of caveolins, the major membrane proteins of caveolae. As well as providing evidence for ubiquitin-mediated endosomal sorting and degradation of caveolin in multivesicular bodies (MVBs), the new findings question the existence of a unique organelle proposed nine years ago, the caveosome.

scholarly journals Regulators of Vps4 ATPase Activity at Endosomes Differentially Influence the Size and Rate of Formation of Intralumenal Vesicles

2010 ◽  
Vol 21 (6) ◽  
pp. 1023-1032 ◽  
Author(s):  
Daniel P. Nickerson ◽  
Matthew West ◽  
Ryan Henry ◽  
Greg Odorizzi
Keyword(s):  
Subcellular Localization ◽  
Atpase Activity ◽  
Transmembrane Proteins ◽  
Regulatory Proteins ◽  
Multivesicular Bodies ◽  
Unique Role ◽  
Vesicle Size ◽  
Endosomal Sorting ◽  
Membrane Scission

Recruitment of endosomal sorting complexes required for transport (ESCRTs) to the cytosolic face of endosomes regulates selective inclusion of transmembrane proteins into the lumenal vesicles of multivesicular bodies (MVBs). ESCRT-0, -I, and -II bind directly to ubiquitinated transmembrane cargoes of the MVB pathway, whereas polymerization of ESCRT-III at endosomes is thought to bend the membrane and/or provide the energetic force that drives membrane scission and detachment of vesicles into the endosome lumen. Disassembly of the ESCRT-III polymer and dissociation of its subunits from endosomes requires the Vps4 ATPase, the activity of which is controlled in vivo by regulatory proteins. We identify distinct spatiotemporal roles for Vps4-regulating proteins through examinations of subcellular localization and endosome morphology. Did2 plays a unique role in the regulation of MVB lumenal vesicle size, whereas Vtal and Vps60 promote efficient membrane scission and delivery of membrane to the endosome lumen. These morphological effects probably result from Vps4-mediated manipulations of ESCRT-III, because we show dissociation of ESCRT-0, -I, and -II from endosomes is not directly dependent on Vps4 activity.

scholarly journals Recruitment of functionally distinct membrane proteins to chromatin mediates nuclear envelope formation in vivo

2009 ◽  
Vol 186 (6) ◽  
pp. 929-929
Author(s):  
Daniel J. Anderson ◽  
Jesse D. Vargas ◽  
Joshua P. Hsiao ◽  
Martin W. Hetzer
Keyword(s):  
Membrane Proteins ◽  
Nuclear Envelope ◽  
Nuclear Envelope Formation ◽  
Distinct Membrane

scholarly journals ESCRT ubiquitin-binding domains function cooperatively during MVB cargo sorting

2009 ◽  
Vol 185 (2) ◽  
pp. 213-224 ◽  
Author(s):  
S. Brookhart Shields ◽  
Andrea J. Oestreich ◽  
Stanley Winistorfer ◽  
Doris Nguyen ◽  
Johanna A. Payne ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Structural Information ◽  
Multivesicular Bodies ◽  
Endosomal Sorting ◽  
Binding Domains ◽  
Ubiquitin Binding ◽  
Cargo Sorting

Ubiquitin (Ub) sorting receptors facilitate the targeting of ubiquitinated membrane proteins into multivesicular bodies (MVBs). Ub-binding domains (UBDs) have been described in several endosomal sorting complexes required for transport (ESCRT). Using available structural information, we have investigated the role of the multiple UBDs within ESCRTs during MVB cargo selection. We found a novel UBD within ESCRT-I and show that it contributes to MVB sorting in concert with the known UBDs within the ESCRT complexes. These experiments reveal an unexpected level of coordination among the ESCRT UBDs, suggesting that they collectively recognize a diverse set of cargo rather than act sequentially at discrete steps.

scholarly journals Ear1p and Ssh4p Are New Adaptors of the Ubiquitin Ligase Rsp5p for Cargo Ubiquitylation and Sorting at Multivesicular Bodies

2008 ◽  
Vol 19 (6) ◽  
pp. 2379-2388 ◽  
Author(s):  
Sébastien Léon ◽  
Zoi Erpapazoglou ◽  
Rosine Haguenauer-Tsapis
Keyword(s):  
Plasma Membrane ◽  
Membrane Proteins ◽  
Ubiquitin Ligase ◽  
Multivesicular Bodies ◽  
Endosomal Sorting ◽  
Frame Fusion ◽  
Vacuolar Targeting

The ubiquitylation of membrane proteins destined for the vacuole/lysosome is essential for their recognition by the endosomal sorting machinery and their internalization into vesicles of multivesicular bodies (MVBs). In yeast, this process requires Rsp5p, an essential ubiquitin ligase of the Nedd4 family. We describe here two redundant proteins, Ear1p and Ssh4p, required for the vacuolar targeting of several cargoes originating from the Golgi or the plasma membrane. Ear1p is an endosomal protein that interacts with Rsp5p through its PPxY motifs, and it is required for the ubiquitylation of selected cargoes before their MVB sorting. In-frame fusion of cargo to ubiquitin overcomes the need for Ear1p/Ssh4p, confirming a role for these proteins in cargo ubiquitylation. Interestingly, Ear1p is itself ubiquitylated by Rsp5p and targeted to the vacuole. Finally, Ear1p overexpression leads to Rsp5p accumulation at endosomes, interfering with some of its functions in trafficking. Therefore, Ear1p/Ssh4p recruit Rsp5p and assist it in its function at MVBs by directing the ubiquitylation of specific cargoes.

scholarly journals Bro1 binding to Snf7 regulates ESCRT-III membrane scission activity in yeast

2011 ◽  
Vol 192 (2) ◽  
pp. 295-306 ◽  
Author(s):  
Megan Wemmer ◽  
Ishara Azmi ◽  
Matthew West ◽  
Brian Davies ◽  
David Katzmann ◽  
...  
Keyword(s):  
Electron Tomography ◽  
Enveloped Viruses ◽  
Endosomal Sorting ◽  
Reduced Frequency ◽  
Stabilization Effect ◽  
Auxiliary Protein ◽  
Membrane Scission ◽  
The Stability

Endosomal sorting complexes required for transport (ESCRTs) promote the invagination of vesicles into the lumen of endosomes, the budding of enveloped viruses, and the separation of cells during cytokinesis. These processes share a topologically similar membrane scission event facilitated by ESCRT-III assembly at the cytosolic surface of the membrane. The Snf7 subunit of ESCRT-III in yeast binds directly to an auxiliary protein, Bro1. Like ESCRT-III, Bro1 is required for the formation of intralumenal vesicles at endosomes, but its role in membrane scission is unknown. We show that overexpression of Bro1 or its N-terminal Bro1 domain that binds Snf7 enhances the stability of ESCRT-III by inhibiting Vps4-mediated disassembly in vivo and in vitro. This stabilization effect correlates with a reduced frequency in the detachment of intralumenal vesicles as observed by electron tomography, implicating Bro1 as a regulator of ESCRT-III disassembly and membrane scission activity.

scholarly journals ENDOSOMAL MEMBRANE TENSION CONTROLS ESCRT-III-DEPENDENT INTRA-LUMENAL VESICLE FORMATION

2019 ◽  
Author(s):  
Vincent Mercier ◽  
Jorge Larios ◽  
Guillaume Molinard ◽  
Antoine Goujon ◽  
Stefan Matile ◽  
...  
Keyword(s):  
Vesicle Formation ◽  
Membrane Tension ◽  
Membrane Remodeling ◽  
Intracellular Traffic ◽  
Physiological Conditions ◽  
Endosomal Membrane ◽  
Endosome Trafficking ◽  
Endosomal Membranes

AbstractPlasma membrane tension strongly affects cell surface processes, such as migration, endocytosis and signalling. However, it is not known whether membrane tension of organelles regulates their functions, notably intracellular traffic. The ESCRT-III complex is the major membrane remodelling complex that drives Intra-Lumenal Vesicle (ILV) formation on endosomal membranes. Here, we made use of a new fluorescent membrane tension probe to show that ESCRT-III subunits are recruited onto endosomal membranes when membrane tension is reduced. We find that tension-dependent recruitment is associated with ESCRT-III polymerization and membrane deformation in vitro, and correlates with increased ILVs formation in ESCRT-III decorated endosomes in vivo. Finally, we find that endosomal membrane tension decreases when ILV formation is triggered by EGF under physiological conditions. These results indicate that membrane tension is a major regulator of ILV formation and of endosome trafficking, leading us to conclude that membrane tension can control organelle functions.One Sentence SummaryMembrane tension decrease facilitates membrane remodeling by ESCRT-III polymerization during intra-lumenal vesicle formation.

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