Membrane trafficking has heretofore been studied with intact organelles. Here, fusion-competent proteoliposomes were reconstituted from a yeast vacuole detergent extract. Homotypic vacuole fusion requires many membrane proteins, including the Ypt7p guanosine triphosphatase and a "SNARE complex" with Vam3p and Nyv1p. Proteoliposomes from extracts immunodepleted of either Vam3p or Ypt7p could not fuse, but vesicles reconstituted from a mixture of these depleted extracts had restored fusion activity. Purified preassembled vacuolar SNARE complex, when reconstituted with a SNARE-depleted extract, was fully functional for fusion. Thus, solubilized integral membrane components can be reconstituted for priming, docking, and fusion steps of organelle trafficking.
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| Gene/Complex | Qualifier | Gene Ontology Term | Annotation Extension | Evidence | Source | Assigned On |
|---|---|---|---|---|---|---|
| SEC17 | involved in | vacuole fusion, non-autophagic | IDA | SGD | 2013-08-07 | |
| SEC18 | involved in | vacuole fusion, non-autophagic | IDA | SGD | 2013-08-07 | |
| NYV1 | involved in | vacuole fusion, non-autophagic | IDA | SGD | 2013-08-07 | |
| VAM3 | involved in | vacuole fusion, non-autophagic | IDA | SGD | 2013-08-07 |
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| Evidence ID | Analyze ID | Interactor | Interactor Systematic Name | Interactor | Interactor Systematic Name | Allele | Assay | Annotation | Action | Phenotype | SGA score | P-value | Source | Reference | Note |
|---|
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| Interactor | Interactor | Assay | Annotation | Action | Modification | |
|---|---|---|---|---|---|---|
| SEC17 | NYV1 | Co-fractionation | manually curated | Bait-Hit | No Modification | |
| SEC17 | SEC18 | Co-fractionation | manually curated | Bait-Hit | No Modification | |
| SEC17 | VAM3 | Co-fractionation | manually curated | Bait-Hit | No Modification | |
| SEC17 | YPT7 | Co-fractionation | manually curated | Bait-Hit | No Modification |