Sphingomyelin organization is required for vesicle biogenesis at the Golgi complex

Durán, Juan M.; Campelo, Felix; Galen, Josse van; Sachsenheimer, Timo; Sot, Jesús; Egorov, Mikhail V.; Rentero, Carles; Enrich, Carlos; Polishchuk, Roman; Goñi, Félix M.; Brügger, Britta; Wieland, Felix; Malhotra, Vivek

doi:10.1038/emboj.2012.317

Cited by 83 publications

(84 citation statements)

References 61 publications

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“…GPI-AP segregation and sorting upon vesicle transport are proposed to be driven by their clustering into special membrane domains or 'lipid rafts' that are enriched in saturated lipids, such as sphingolipids and sterols, which would act as selective platforms for vesicle budding . This model is supported by the observations that purified Golgi-derived vesicles in yeast are enriched in sphingolipids and ergosterol (Klemm et al, 2009), and that long-chain sphingomyelin is required for vesicle biogenesis at the Golgi complex in mammalian cells (Duran et al, 2012).…”

Section: Introductionsupporting

confidence: 67%

Sorting of GPI-anchored proteins from yeast to mammals – common pathways at different sites?

Muñiz

Zurzolo

2014

Journal of Cell Science

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Glycosylphosphatidylinositol (GPI)-anchored proteins (GPI-APs) are luminal secretory cargos that are attached by a post-translational glycolipid modification, the GPI anchor, to the external leaflet of the plasma membrane. GPI-APs are conserved among eukaryotes and possess many diverse and vital functions for which the GPI membrane attachment appears to be essential. The presence of the GPI anchor and its subsequent modifications along the secretory pathway confer to the anchored proteins unique trafficking properties that make GPIAPs an exceptional system to study mechanisms of sorting. In this Commentary, we discuss the recent advances in the field of GPI-AP sorting focusing on the mechanisms operating at the level of the exit from the ER and from the trans-Golgi network (TGN), which take place, respectively, in yeast and in polarized mammalian cells. By considering the similarities and differences between these two sorting events, we present unifying principles that appear to work at different sorting stations and in different organisms.

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Section: Introductionsupporting

confidence: 67%

Sorting of GPI-anchored proteins from yeast to mammals – common pathways at different sites?

Muñiz

Zurzolo

2014

Journal of Cell Science

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“…An appealingly simple hypothesis is that the more voluminous exoplasmic hemiTMDs of Golgi proteins negatively affect their fit into the highly curved membrane domains from which Golgi transport carriers enriched in cholesterol and sphingolipids originate (Duran et al, 2012;Klemm et al, 2009;Polishchuk et al, 2003). By contrast, TMDs from plasma membrane proteins could conform a complex, thermodynamically favourable association with these order-inducing lipids and exit the Golgi.…”

Section: Features Of the Exoplasmic Hemi-tmds Define Golgi Or Plasmamentioning

confidence: 99%

Short length transmembrane domains having voluminous exoplasmic halves determine retention of Type II membrane proteins in the Golgi complex

Quiroga¹,

Trenchi²,

Montoro³

et al. 2013

Journal of Cell Science

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SummaryIt is still unclear why some proteins that travel along the secretory pathway are retained in the Golgi complex whereas others make their way to the plasma membrane. Recent bioinformatic analyses on a large number of single-spanning membrane proteins support the hypothesis that specific features of the transmembrane domain (TMD) are relevant to the sorting of these proteins to particular organelles. Here we experimentally test this hypothesis for Golgi and plasma membrane proteins. Using the Golgi SNARE protein Sft1 and the plasma membrane SNARE protein Sso1 from Saccharomyces cerevisiae as model proteins, we modified the length of their TMDs and the volume of their exoplasmic hemi-TMD, and determined their subcellular localization both in yeast and mammalian cells. We found that short TMDs with high-volume exoplasmic hemi-TMDs confer Golgi membrane residence, whereas TMDs with lowvolume exoplasmic hemi-TMDs, either short or long, confer plasma membrane residence to these proteins. Results indicate that the shape of the exoplasmic hemi-TMD, in addition to the length of the entire TMD, determine retention in the Golgi or exit to the plasma membrane of Type II membrane proteins.

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“…This postulate is supported by results obtained with D-ceramide-C6. This lipid derivative causes the disruption of cholesterol-and sphingomyelin-enriched domains at the Golgi (63), which is accompanied by a swelling of cisternae and alterations in N-glycosylation (63,66), both strikingly coincidental with impairments at the Golgi occurring after actin depolymerization and pharmacological V-ATPase inhibition (49,57,58). Importantly, D-ceramide-C6 treatment also caused Golgi and TGN pH alkalization, which also occurs after actin-depolymerizing toxins (49).…”

Section: Table 2 Reduction Of Golgi Membrane Lipid Order Increases Gomentioning

confidence: 99%

“…Thus, we exogenously added short chain ceramide-C6, the non-metabolizable enantiomer L-ceramide-C6 (N-hexanoyl-L-erythrosphingosine) as a control, and the enantiomer D-ceramide-C6 (N-hexanoyl-D-erythrosphingosine), which is incorporated into Golgi membranes and locally increases levels of shortchain sphingomyelin, short-chain glucosylceramide, and diacylglycerol. As a consequence, there is a reduction in the formation of cholesterol/sphingomyelin-enriched domains into the Golgi (63). Thereafter, we measured Golgi and TGN pH in cells transfected with Golgi-or TGN-pHluorin constructs, and we subsequently cells treated with D-or L-ceramide-C6 (20 M for 30 min each).…”

Section: V-atpase Revealed By Gfp-tagged Subunit B2 Of the V 1 Domainmentioning

confidence: 99%

Actin Filaments Are Involved in the Coupling of V0-V1 Domains of Vacuolar H+-ATPase at the Golgi Complex

Serra‐Peinado¹,

Sicart²,

Llopis³

et al. 2016

Journal of Biological Chemistry

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We previously reported that actin-depolymerizing agents promote the alkalization of the Golgi stack and the trans-Golgi network. The main determinant of acidic pH at the Golgi is the vacuolar-type H ؉ -translocating ATPase (V-ATPase), whose V 1 domain subunits B and C bind actin. We have generated a GFPtagged subunit B2 construct (GFP-B2) that is incorporated into the V 1 domain, which in turn is coupled to the V 0 sector. GFP-B2 subunit is enriched at distal Golgi compartments in HeLa cells. Subcellular fractionation, immunoprecipitation, and inversal FRAP experiments show that the actin depolymerization promotes the dissociation of V 1 -V 0 domains, which entails subunit B2 translocation from Golgi membranes to the cytosol. Moreover, molecular interaction between subunits B2 and C1 and actin were detected. In addition, Golgi membrane lipid order disruption by D-ceramide-C6 causes Golgi pH alkalization. We conclude that actin regulates the Golgi pH homeostasis maintaining the coupling of V 1 -V 0 domains of V-ATPase through the binding of microfilaments to subunits B and C and preserving the integrity of detergent-resistant membrane organization. These results establish the Golgi-associated V-ATPase activity as the molecular link between actin and the Golgi pH.The secretory pathway is characterized by progressive lumen acidification of its organelles, from almost neutral in the endoplasmic reticulum (ER) 4 (pH Ϸ7.1-7.2), along the cis-to-trans Golgi stack (pH Ϸ6.7-6.0), to more acidic in the trans-Golgi network (TGN) and secretory vesicles/granules (pH Ϸ5.0) (1-3). This pH gradient is crucial for post-translational modifications and membrane trafficking events (4, 5). The main molecular determinant of the progressive fall in pH along the secretory pathway is the vacuolar [H ϩ ]ATPase (V-ATPase) (6 -8). V-ATPase is a multisubunit complex composed of two large domains, V 0 and V 1 . The V 0 domain is a 260-kDa integral membrane complex made up of five different subunits (a, b, c, cЈ, cЉ, d, and e), which mediates proton translocation; the V 1 domain is a 600 -650-kDa peripheral complex composed of eight different subunits (A, B, C, D, E, F, G, and H), which is responsible for the ATP hydrolysis that provides the mechanical force necessary for proton (H ϩ ) translocation (7, 9 -11). Whereas they are the primary source of proton delivery to endomembranes consuming ATP, the final steady-state pH in the secretory pathway is the result of the balance between active H ϩ pumping by the V-ATPase, passive H ϩ efflux through organelle endogenous H ϩ permeability, and differences in counter-ion conductance (3, 12).How differences in the pH of individual secretory compartments are generated is not well understood. Differential VATPase density and/or local regulatory mechanisms in secretory organelles and subcompartments are possible (13). In this respect, V-ATPase-dependent proton translocation could be regulated by several mechanisms, which include the following: (a) differential V-ATPase subunit expression; (b) intracel...

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Sphingomyelin organization is required for vesicle biogenesis at the Golgi complex

Cited by 83 publications

References 61 publications

Sorting of GPI-anchored proteins from yeast to mammals – common pathways at different sites?

Sorting of GPI-anchored proteins from yeast to mammals – common pathways at different sites?

Short length transmembrane domains having voluminous exoplasmic halves determine retention of Type II membrane proteins in the Golgi complex

Actin Filaments Are Involved in the Coupling of V0-V1 Domains of Vacuolar H+-ATPase at the Golgi Complex

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