Autophagy is an ubiquitous degradative process in eukaryotic cells. It is involved in various developmental programs and is also implicated in human pathophysiology. The basic process involves the formation of a cytosolic double membrane vesicle, termed an autophagosome. The autophagosome sequesters bulk cytosol, and after completion its outside membrane fuses with the limiting membrane of the lysosome/vacuole. The fusion event releases the inner single-membrane vesicle into the lysosome/vacuole lumen where the vesicle is now termed an autophagic body. The autophagic body and its cargo are typically degraded, and the resulting micromolecules are recycled for subsequent use in the cytosol. Autophagy is the only pathway with the capacity to degrade entire organelles. Accordingly, it may play a critical role in preventing pathologies that results from damaged organelles including the mitochondria, or from the accumulation of large protein aggregates, such as occur in certain types of neurodegenerative diseases. This article describes an assay to monitor bulk autophagy in yeast. The marker protein is a cytosolic derivative of the vacuolar enzyme alkaline phosphatase, Pho8. Following uptake into the vacuole, the precursor enzyme is cleaved at the C terminus to generate the active form. Cells expressing the cytosolic form of Pho8, Pho8Delta60, are assayed for alkaline phosphatase activity before and after shifting to conditions that induce autophagy.
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Evidence ID | Analyze ID | Gene/Complex | Systematic Name/Complex Accession | Qualifier | Gene Ontology Term ID | Gene Ontology Term | Aspect | Annotation Extension | Evidence | Method | Source | Assigned On | Reference |
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Evidence ID | Analyze ID | Gene | Gene Systematic Name | Phenotype | Experiment Type | Experiment Type Category | Mutant Information | Strain Background | Chemical | Details | Reference |
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Evidence ID | Analyze ID | Gene | Gene Systematic Name | Disease Ontology Term | Disease Ontology Term ID | Qualifier | Evidence | Method | Source | Assigned On | Reference |
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Evidence ID | Analyze ID | Regulator | Regulator Systematic Name | Target | Target Systematic Name | Direction | Regulation of | Happens During | Regulator Type | Direction | Regulation Of | Happens During | Method | Evidence | Strain Background | Reference |
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Site | Modification | Modifier | Source | Reference |
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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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Evidence ID | Analyze ID | Interactor | Interactor Systematic Name | Interactor | Interactor Systematic Name | Assay | Annotation | Action | Modification | Source | Reference | Note |
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Complement ID | Locus ID | Gene | Species | Gene ID | Strain background | Direction | Details | Source | Reference |
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Evidence ID | Analyze ID | Dataset | Description | Keywords | Number of Conditions | Reference |
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