Vps35 (CG5625) is the Drosophila ortholog of VPS35, the large alpha-solenoid backbone subunit of the retromer cargo-selective complex, which binds Vps26 at its N-terminal region and Vps29 at its C-terminal region. Recruited to endosomes by Rab GTPases and sorting nexins, the Vps35-Vps26-Vps29 trimer recognizes transmembrane cargo and retrieves it to the trans-Golgi network or recycles it to the plasma membrane, sparing it from lysosomal degradation. In flies Vps35 is required to retrieve the Wnt cargo receptor Wntless to the Golgi and hence for Wingless (but not Hedgehog) secretion, to recycle the luminal protein Serpentine via Rab9-positive late endosomes in the trachea, and to retrieve Notch in neural progenitors. Vps35 loss also impairs endocytosis, synaptic vesicle recycling at the neuromuscular junction and dopaminergic transmission, and it interacts genetically with the Parkinson's disease kinase LRRK2. At larval neuromuscular junctions Vps35 is present pre- and postsynaptically and limits loading of APP-like cargo into extracellular vesicles.
| GO Term | Evidence | Action | Reason |
|---|---|---|---|
| GO:0005515 protein binding | IPI PMID:29909971 Phospholipase PLA2G6, a Parkinsonism-Associated Gene, Affect... | MODIFY | Summary: Binding to the phospholipase iPLA2-VIA (FBgn0036053). Reason: The IPI records Vps35 binding to iPLA2-VIA, the fly PLA2G6 homolog, which enhances retromer function. phospholipase binding is the informative term. Proposed replacements: phospholipase binding Supporting Evidence: PMID:29909971 iPLA2-VIA binds the retromer subunits Vps35 and Vps26 and enhances retromer function to promote protein and lipid recycling. |
| GO:0005768 endosome | IDA PMID:18057029 Drosophila Vps35 function is necessary for normal endocytic ... | ACCEPT | Summary: Retromer acts on endosomes. Reason: Correct location; early and late endosome terms are also annotated by IDA. Supporting Evidence: PMID:23322046 Vps35, WASH and actin filaments differentially localize at the Rab9-enriched subdomains of the endosomal membrane |
| GO:0005768 endosome | ISS GO_REF:0000024 | ACCEPT | Summary: Retromer acts on endosomes. Reason: Correct location; early and late endosome terms are also annotated by IDA. Supporting Evidence: PMID:23322046 Vps35, WASH and actin filaments differentially localize at the Rab9-enriched subdomains of the endosomal membrane |
| GO:0005769 early endosome | IDA PMID:22041890 SNX3 controls Wingless/Wnt secretion through regulating retr... | ACCEPT | Summary: Vps35 colocalizes with Snx3 on early endosomes. Reason: Direct localization in fly cells. Supporting Evidence: PMID:22041890 DSNX3 can interact with the retromer component Vps35, and co-localize with Vps35 in early endosomes |
| GO:0005770 late endosome | IBA GO_REF:0000033 | ACCEPT | Summary: Vps35 localizes to Rab9-positive late endosome domains. Reason: Direct localization in S2 cells and tracheal cells (IDA); IBA donors include fly Vps35 and Arabidopsis VPS35. Supporting Evidence: PMID:23322046 Vps35 and Serp colocalize at late endosomal Rab9 domains |
| GO:0005770 late endosome | IDA PMID:23322046 Rab9 and retromer regulate retrograde trafficking of luminal... | ACCEPT | Summary: Vps35 localizes to Rab9-positive late endosome domains. Reason: Direct localization in S2 cells and tracheal cells (IDA); IBA donors include fly Vps35 and Arabidopsis VPS35. Supporting Evidence: PMID:23322046 Vps35 and Serp colocalize at late endosomal Rab9 domains |
| GO:0005829 cytosol | IEA GO_REF:0000108 | KEEP AS NON CORE | Summary: Cytosolic pool of a peripheral membrane complex. Reason: Retromer cycles between cytosol and endosome membranes; the functional location is the endosome. Inferred from the process annotation (GO_REF:0000108). |
| GO:0006897 endocytosis | IMP PMID:18057029 Drosophila Vps35 function is necessary for normal endocytic ... | KEEP AS NON CORE | Summary: Vps35 RNAi inhibits scavenger-receptor ligand uptake in S2 cells. Reason: Loss of Vps35 mislocalizes receptors and endocytic proteins and inhibits endocytosis, likely indirectly through failed receptor recycling and altered actin; endocytosis is as specific as the evidence allows but peripheral to retromer's core role. Supporting Evidence: PMID:18057029 Loss of Vps35 inhibits scavenger receptor ligand endocytosis, and causes mislocalisation of a number of receptors and endocytic proteins. |
| GO:0012505 endomembrane system | NAS PMID:26172538 Retromer Ensures the Degradation of Autophagic Cargo by Main... | MODIFY | Summary: Generic endomembrane system; less specific than endosome. Reason: Correct but less specific than the evidence supports; endosome (GO:0005768) is already annotated to Vps35 (IDA, ISS). Proposed replacements: endosome Supporting Evidence: PMID:23322046 Vps35, WASH and actin filaments differentially localize at the Rab9-enriched subdomains of the endosomal membrane |
| GO:0015031 protein transport | IEA GO_REF:0000002 | MODIFY | Summary: Protein transport; correct but less specific than endosome-to-Golgi retrieval. Reason: The more specific process retrograde transport, endosome to Golgi (GO:0042147) is already annotated to Vps35 (IMP, IBA, ISS, IEA, NAS). IEA from InterPro IPR005378. Proposed replacements: retrograde transport, endosome to Golgi Supporting Evidence: PMID:18193037 Following dissociation from Wingless, Wntless is internalized and returns to the Golgi apparatus in a retromer-dependent manner. |
| GO:0030904 retromer complex | IBA GO_REF:0000033 | MODIFY | Summary: Retromer complex; correct but less specific than the cargo-selective subcomplex. Reason: Vps35 is the backbone of the Vps35-Vps26-Vps29 cargo-selective trimer of retromer; retromer, cargo-selective complex (GO:0030906) is already annotated to Vps35 (ISS, IEA). Proposed replacements: retromer, cargo-selective complex Supporting Evidence: PMID:32286230 Retromer, including Vps35, Vps26, and Vps29, is a protein complex responsible for recycling proteins within the endolysosomal pathway. |
| GO:0030904 retromer complex | NAS PMID:26172538 Retromer Ensures the Degradation of Autophagic Cargo by Main... | MODIFY | Summary: Retromer complex; correct but less specific than the cargo-selective subcomplex. Reason: Vps35 is the backbone of the Vps35-Vps26-Vps29 cargo-selective trimer of retromer; retromer, cargo-selective complex (GO:0030906) is already annotated to Vps35 (ISS, IEA). Proposed replacements: retromer, cargo-selective complex Supporting Evidence: PMID:32286230 Retromer, including Vps35, Vps26, and Vps29, is a protein complex responsible for recycling proteins within the endolysosomal pathway. |
| GO:0030906 retromer, cargo-selective complex | IEA GO_REF:0000002 | ACCEPT | Summary: Vps35 is the core subunit of the retromer cargo-selective complex. Reason: Vps35 assembles with Vps26 and Vps29 into the cargo-selective trimer (ISS from yeast VPS35; IEA from InterPro). Supporting Evidence: PMID:32286230 Vps35 and Vps26 proteins are normally expressed and associated |
| GO:0030906 retromer, cargo-selective complex | ISS GO_REF:0000024 | ACCEPT | Summary: Vps35 is the core subunit of the retromer cargo-selective complex. Reason: Vps35 assembles with Vps26 and Vps29 into the cargo-selective trimer (ISS from yeast VPS35; IEA from InterPro). Supporting Evidence: PMID:32286230 Vps35 and Vps26 proteins are normally expressed and associated |
| GO:0031267 small GTPase binding | IDA PMID:28482024 Vps35 in cooperation with LRRK2 regulates synaptic vesicle e... | ACCEPT | Summary: Small GTPase binding reported in a Vps35-LRRK2 synaptic study. Reason: Only the abstract is cached; the curator's IDA is accepted, consistent with the independently shown Rab9 binding (PMID:23322046) and the involvement of Rab5/Rab11 in the Vps35 synaptic phenotype. Supporting Evidence: PMID:28482024 Drosophila LRRK2 dLRRK together with Rab5 and Rab11 is also implicated in synaptic vesicle recycling |
| GO:0031267 small GTPase binding | IPI PMID:23322046 Rab9 and retromer regulate retrograde trafficking of luminal... | ACCEPT | Summary: Vps35 binds Rab9 in S2 cell lysates. Reason: GST-Rab9 pulled down myc-Vps35; Rab GTPases (Rab7, Rab9) recruit and regulate retromer on late endosomes, so small GTPase binding is a mechanistically relevant activity. Supporting Evidence: PMID:23322046 both GTP- and GDP-loaded forms of GST-Rab9, but not GST pulled down myc-Vps35 |
| GO:0031594 neuromuscular junction | IDA PMID:28482024 Vps35 in cooperation with LRRK2 regulates synaptic vesicle e... | KEEP AS NON CORE | Summary: Vps35 at fly synaptic terminals. Reason: Synaptic localizations from the Vps35-LRRK2 study (only the abstract is cached; curator's IDA accepted). Valid, specific sites of action in neurons but peripheral to the core endosomal location. Supporting Evidence: PMID:28482024 the loss of Drosophila Vps35 (dVps35) affects synaptic vesicle recycling, dopaminergic synaptic release |
| GO:0031594 neuromuscular junction | IMP PMID:28482024 Vps35 in cooperation with LRRK2 regulates synaptic vesicle e... | KEEP AS NON CORE | Summary: Vps35 at fly synaptic terminals. Reason: Synaptic localizations from the Vps35-LRRK2 study (only the abstract is cached; curator's IDA accepted). Valid, specific sites of action in neurons but peripheral to the core endosomal location. Supporting Evidence: PMID:28482024 the loss of Drosophila Vps35 (dVps35) affects synaptic vesicle recycling, dopaminergic synaptic release |
| GO:0032456 endocytic recycling | NAS PMID:26172538 Retromer Ensures the Degradation of Autophagic Cargo by Main... | ACCEPT | Summary: Retromer recycles cargo from endosomes back to the plasma membrane. Reason: Besides TGN retrieval, Vps35-retromer recycles cargo such as Notch to the cell surface in fly neural progenitors. Supporting Evidence: PMID:30176986 Retromer (purple) normally interacts with Notch (green) and retrieves the pool of hypoubiquitinated Notch evading the ESCRT (cyan)-lysosomal degradation pathway and sends it back to the cell surface |
| GO:0038024 cargo receptor activity | IPI PMID:30176986 The retromer complex safeguards against neural progenitor-de... | MODIFY | Summary: Retromer binds Notch as cargo; retromer acts as a cytosolic cargo adaptor, not a transmembrane cargo receptor. Reason: Notch co-immunoprecipitates with Vps35 from larval brain and is retrieved by retromer, so Notch is a bona fide retromer cargo. GO cargo receptor activity is defined for membrane-spanning receptors; the activity of the peripheral Vps35-Vps26-Vps29 trimer is cargo adaptor activity (already annotated to Vps35 by IBA), contributed jointly by the subunits. Proposed replacements: cargo adaptor activity Supporting Evidence: PMID:30176986 Notch-V5 expressed in central brain neuroblast lineages was specifically coimmunoprecipitated with Vps35-FLAG from fly larval brain extracts |
| GO:0042147 retrograde transport, endosome to Golgi | IBA GO_REF:0000033 | ACCEPT | Summary: Core retromer process; Vps35 retrieves Wntless and Serpentine to the Golgi. Reason: Vps35 mutant cells fail to retrieve Wntless to the Golgi, and Vps35 is required for retrograde trafficking of endocytosed Serpentine from late endosomes to the TGN in the trachea. Supporting Evidence: PMID:18193037 Following dissociation from Wingless, Wntless is internalized and returns to the Golgi apparatus in a retromer-dependent manner. PMID:23322046 The requirement for Rab9 and Vps35 for the luminal retention of Serp suggests that the endocytosed Serp is sorted from LEs to the TGN for secretion into the lumen |
| GO:0042147 retrograde transport, endosome to Golgi | IEA GO_REF:0000002 | ACCEPT | Summary: Core retromer process; Vps35 retrieves Wntless and Serpentine to the Golgi. Reason: Vps35 mutant cells fail to retrieve Wntless to the Golgi, and Vps35 is required for retrograde trafficking of endocytosed Serpentine from late endosomes to the TGN in the trachea. Supporting Evidence: PMID:18193037 Following dissociation from Wingless, Wntless is internalized and returns to the Golgi apparatus in a retromer-dependent manner. PMID:23322046 The requirement for Rab9 and Vps35 for the luminal retention of Serp suggests that the endocytosed Serp is sorted from LEs to the TGN for secretion into the lumen |
| GO:0042147 retrograde transport, endosome to Golgi | IMP PMID:23322046 Rab9 and retromer regulate retrograde trafficking of luminal... | ACCEPT | Summary: Core retromer process; Vps35 retrieves Wntless and Serpentine to the Golgi. Reason: Vps35 mutant cells fail to retrieve Wntless to the Golgi, and Vps35 is required for retrograde trafficking of endocytosed Serpentine from late endosomes to the TGN in the trachea. Supporting Evidence: PMID:18193037 Following dissociation from Wingless, Wntless is internalized and returns to the Golgi apparatus in a retromer-dependent manner. PMID:23322046 The requirement for Rab9 and Vps35 for the luminal retention of Serp suggests that the endocytosed Serp is sorted from LEs to the TGN for secretion into the lumen |
| GO:0042147 retrograde transport, endosome to Golgi | ISS GO_REF:0000024 | ACCEPT | Summary: Core retromer process; Vps35 retrieves Wntless and Serpentine to the Golgi. Reason: Vps35 mutant cells fail to retrieve Wntless to the Golgi, and Vps35 is required for retrograde trafficking of endocytosed Serpentine from late endosomes to the TGN in the trachea. Supporting Evidence: PMID:18193037 Following dissociation from Wingless, Wntless is internalized and returns to the Golgi apparatus in a retromer-dependent manner. PMID:23322046 The requirement for Rab9 and Vps35 for the luminal retention of Serp suggests that the endocytosed Serp is sorted from LEs to the TGN for secretion into the lumen |
| GO:0042147 retrograde transport, endosome to Golgi | NAS PMID:26172538 Retromer Ensures the Degradation of Autophagic Cargo by Main... | ACCEPT | Summary: Core retromer process; Vps35 retrieves Wntless and Serpentine to the Golgi. Reason: Vps35 mutant cells fail to retrieve Wntless to the Golgi, and Vps35 is required for retrograde trafficking of endocytosed Serpentine from late endosomes to the TGN in the trachea. Supporting Evidence: PMID:18193037 Following dissociation from Wingless, Wntless is internalized and returns to the Golgi apparatus in a retromer-dependent manner. PMID:23322046 The requirement for Rab9 and Vps35 for the luminal retention of Serp suggests that the endocytosed Serp is sorted from LEs to the TGN for secretion into the lumen |
| GO:0043195 terminal bouton | IDA PMID:28482024 Vps35 in cooperation with LRRK2 regulates synaptic vesicle e... | KEEP AS NON CORE | Summary: Vps35 at fly synaptic terminals. Reason: Synaptic localizations from the Vps35-LRRK2 study (only the abstract is cached; curator's IDA accepted). Valid, specific sites of action in neurons but peripheral to the core endosomal location. Supporting Evidence: PMID:28482024 the loss of Drosophila Vps35 (dVps35) affects synaptic vesicle recycling, dopaminergic synaptic release |
| GO:0048488 synaptic vesicle endocytosis | IMP PMID:28482024 Vps35 in cooperation with LRRK2 regulates synaptic vesicle e... | KEEP AS NON CORE | Summary: Vps35 loss affects synaptic vesicle recycling at fly synapses. Reason: Vps35 mutants have synaptic vesicle recycling and dopaminergic release defects, rescued by wild-type but not the PD-associated D647N Vps35. A real but tissue-specific requirement, peripheral to the core endosomal retrieval function (only the abstract is cached). Supporting Evidence: PMID:28482024 the loss of Drosophila Vps35 (dVps35) affects synaptic vesicle recycling, dopaminergic synaptic release and sleep behavior associated with dopaminergic activity |
| GO:0048786 presynaptic active zone | IDA PMID:28482024 Vps35 in cooperation with LRRK2 regulates synaptic vesicle e... | KEEP AS NON CORE | Summary: Vps35 at fly synaptic terminals. Reason: Synaptic localizations from the Vps35-LRRK2 study (only the abstract is cached; curator's IDA accepted). Valid, specific sites of action in neurons but peripheral to the core endosomal location. Supporting Evidence: PMID:28482024 the loss of Drosophila Vps35 (dVps35) affects synaptic vesicle recycling, dopaminergic synaptic release |
| GO:0061357 positive regulation of Wnt protein secretion | IMP PMID:18193037 Wingless secretion requires endosome-to-Golgi retrieval of W... | KEEP AS NON CORE | Summary: Vps35 is required for Wingless secretion through Wntless retrieval. Reason: Wg secretion requires retromer-dependent recycling of its cargo receptor Wntless; the effect on Wg secretion is real and specific (Hh secretion is unaffected) but is a downstream consequence of the core cargo-retrieval activity. Supporting Evidence: PMID:18193037 Without the retromer-dependent recycling route, Wingless secretion is impaired |
| GO:0071212 subsynaptic reticulum | IDA PMID:28482024 Vps35 in cooperation with LRRK2 regulates synaptic vesicle e... | KEEP AS NON CORE | Summary: Vps35 at fly synaptic terminals. Reason: Synaptic localizations from the Vps35-LRRK2 study (only the abstract is cached; curator's IDA accepted). Valid, specific sites of action in neurons but peripheral to the core endosomal location. Supporting Evidence: PMID:28482024 the loss of Drosophila Vps35 (dVps35) affects synaptic vesicle recycling, dopaminergic synaptic release |
| GO:0098691 dopaminergic synapse | IDA PMID:28482024 Vps35 in cooperation with LRRK2 regulates synaptic vesicle e... | KEEP AS NON CORE | Summary: Vps35 at fly synaptic terminals. Reason: Synaptic localizations from the Vps35-LRRK2 study (only the abstract is cached; curator's IDA accepted). Valid, specific sites of action in neurons but peripheral to the core endosomal location. Supporting Evidence: PMID:28482024 the loss of Drosophila Vps35 (dVps35) affects synaptic vesicle recycling, dopaminergic synaptic release |
| GO:0098691 dopaminergic synapse | IMP PMID:28482024 Vps35 in cooperation with LRRK2 regulates synaptic vesicle e... | KEEP AS NON CORE | Summary: Vps35 at fly synaptic terminals. Reason: Synaptic localizations from the Vps35-LRRK2 study (only the abstract is cached; curator's IDA accepted). Valid, specific sites of action in neurons but peripheral to the core endosomal location. Supporting Evidence: PMID:28482024 the loss of Drosophila Vps35 (dVps35) affects synaptic vesicle recycling, dopaminergic synaptic release |
| GO:0098833 presynaptic endocytic zone | IDA PMID:28482024 Vps35 in cooperation with LRRK2 regulates synaptic vesicle e... | KEEP AS NON CORE | Summary: Vps35 at fly synaptic terminals. Reason: Synaptic localizations from the Vps35-LRRK2 study (only the abstract is cached; curator's IDA accepted). Valid, specific sites of action in neurons but peripheral to the core endosomal location. Supporting Evidence: PMID:28482024 the loss of Drosophila Vps35 (dVps35) affects synaptic vesicle recycling, dopaminergic synaptic release |
| GO:0099638 obsolete endosome to plasma membrane protein transport | IBA GO_REF:0000033 | MODIFY | Summary: Obsolete term; retromer returns cargo such as Notch from endosomes to the cell surface. Reason: GO:0099638 is obsolete and was merged into endocytic recycling (GO:0032456). The IMP study models retromer retrieving hypo-ubiquitinated Notch from endosomes to the cell surface; the IBA row (donors include fly Vps35 and mouse Vps35) carries the same meaning. Proposed replacements: endocytic recycling Supporting Evidence: PMID:30176986 Retromer (purple) normally interacts with Notch (green) and retrieves the pool of hypoubiquitinated Notch evading the ESCRT (cyan)-lysosomal degradation pathway and sends it back to the cell surface |
| GO:0099638 obsolete endosome to plasma membrane protein transport | IMP PMID:30176986 The retromer complex safeguards against neural progenitor-de... | MODIFY | Summary: Obsolete term; retromer returns cargo such as Notch from endosomes to the cell surface. Reason: GO:0099638 is obsolete and was merged into endocytic recycling (GO:0032456). The IMP study models retromer retrieving hypo-ubiquitinated Notch from endosomes to the cell surface; the IBA row (donors include fly Vps35 and mouse Vps35) carries the same meaning. Proposed replacements: endocytic recycling Supporting Evidence: PMID:30176986 Retromer (purple) normally interacts with Notch (green) and retrieves the pool of hypoubiquitinated Notch evading the ESCRT (cyan)-lysosomal degradation pathway and sends it back to the cell surface |
| GO:0140312 cargo adaptor activity | IBA GO_REF:0000033 | ACCEPT | Summary: Vps35 contributes to the cargo-adaptor activity of the retromer trimer. Reason: The cargo-selective complex binds cargo such as Wntless and Notch and links it to the sorting-nexin coat; Vps35 is the central cargo-binding scaffold (contributes_to). IBA donor yeast VPS35. Supporting Evidence: PMID:18193037 Furthermore, biochemical evidence confirms that Wntless engages with the retromer complex. PMID:30176986 Notch-V5 expressed in central brain neuroblast lineages was specifically coimmunoprecipitated with Vps35-FLAG from fly larval brain extracts |
| GO:1900242 regulation of synaptic vesicle endocytosis | IDA PMID:28482024 Vps35 in cooperation with LRRK2 regulates synaptic vesicle e... | KEEP AS NON CORE | Summary: Vps35 loss affects synaptic vesicle recycling at fly synapses. Reason: Vps35 mutants have synaptic vesicle recycling and dopaminergic release defects, rescued by wild-type but not the PD-associated D647N Vps35. A real but tissue-specific requirement, peripheral to the core endosomal retrieval function (only the abstract is cached). Supporting Evidence: PMID:28482024 the loss of Drosophila Vps35 (dVps35) affects synaptic vesicle recycling, dopaminergic synaptic release and sleep behavior associated with dopaminergic activity |
| GO:1900242 regulation of synaptic vesicle endocytosis | IMP PMID:28482024 Vps35 in cooperation with LRRK2 regulates synaptic vesicle e... | KEEP AS NON CORE | Summary: Vps35 loss affects synaptic vesicle recycling at fly synapses. Reason: Vps35 mutants have synaptic vesicle recycling and dopaminergic release defects, rescued by wild-type but not the PD-associated D647N Vps35. A real but tissue-specific requirement, peripheral to the core endosomal retrieval function (only the abstract is cached). Supporting Evidence: PMID:28482024 the loss of Drosophila Vps35 (dVps35) affects synaptic vesicle recycling, dopaminergic synaptic release and sleep behavior associated with dopaminergic activity |
| GO:1904647 response to rotenone | IMP PMID:24915984 Vacuolar protein sorting 35 (Vps35) rescues locomotor defici... | MARK AS OVER ANNOTATED | Summary: Vps35 overexpression protects flies from rotenone toxicity. Reason: The evidence is a protective effect of Vps35 overexpression in a parkinsonism model, i.e. a genetic modifier phenotype; it does not show that Vps35 normally participates in a response to rotenone. Supporting Evidence: PMID:24915984 overexpressing Vps35 alone protects from toxicity of rotenone |
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Download this section (compressed HTML)Q: Does the PD-associated Vps35 D647N mutation impair WASH (FAM21) binding in flies as in mammals, and does this explain its failure to rescue synaptic vesicle recycling?
Experiment: Express Vps35 variants defective in FAM21/WASH binding or Vps26 binding in Vps35 null neurons and measure Wntless retrieval, Notch trafficking and FM1-43 uptake.
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Download this section (compressed HTML)Vacuolar protein sorting 35 (Vps35) encodes a component of the retromer complex that acts to recycle membrane proteins to the Golgi or plasma membrane from endocytosed vesicles.
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