The Vacuole Import and Degradation Pathway Utilizes Early Steps of Endocytosis and Actin Polymerization to Deliver Cargo Proteins to the Vacuole for Degradation

• Published in: The Journal of biological chemistry 2010-01, Vol.285 (2), p.1516-1528
• Main Authors: Brown, C. Randell, Dunton, Danielle, Chiang, Hui-Ling
• Format: Article
• Language: English
• Subjects: Actins - genetics; Actins - metabolism; Cell Membrane - genetics; Cell Membrane - metabolism; Cell/Endocytosis; Coatomer Protein - genetics; Coatomer Protein - metabolism; Endocytosis - drug effects; Endocytosis - physiology; FBPase; Fructose-Bisphosphatase - genetics; Fructose-Bisphosphatase - metabolism; Glucose - pharmacology; Malate Dehydrogenase - genetics; Malate Dehydrogenase - metabolism; Organisms/Yeast; Protein Synthesis, Post-Translational Modification, and Degradation; Protein Transport - drug effects; Protein Transport - physiology; Protein/Turnover; Qa-SNARE Proteins; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - metabolism; Saccharomyces cerevisiae Proteins - genetics; Saccharomyces cerevisiae Proteins - metabolism; Subcellular Organelles/Cytoskeleton; Subcellular Organelles/Lysosomes; Subcellular Organelles/Vesicles; Sweetening Agents - pharmacology; Vacuoles - genetics; Vacuoles - metabolism; Vesicular Transport Proteins - genetics; Vesicular Transport Proteins - metabolism; Vid Vesicles
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M109.028241

When glucose is added to yeast cells that are starved for 3 days, fructose-1,6-bisphosphatase (FBPase) and malate dehydrogenase 2 are degraded in the vacuole via the vacuole import and degradation (Vid) pathway. In this study, we examined the distribution of FBPase at the ultrastructural level. FBPase was observed in areas close to the plasma membrane and in cytoplasmic structures that are heterogeneous in size and density. We have isolated these intracellular structures that contain FBPase, the Vid vesicle marker Vid24p, and the endosomal marker Pep12p. They appeared irregular in size and shape. In yeast, actin polymerization plays an important role in early steps of endocytosis. Mutants that affect actin polymerization inhibited FBPase degradation, suggesting that actin polymerization is important for FBPase degradation. Both FBPase and malate dehydrogenase 2 were associated with actin patches. Vid vesicle proteins such as Vid24p or Sec28p were also at actin patches, although they dissociated from these structures at later time points. We propose that Vid24p and Sec28p are present at actin patches during glucose starvation. Cargo proteins arrive at these sites following the addition of glucose, and the endocytic vesicles then pinch off from the plasma membrane. Following the fusion of endosomes with the vacuole, cargo proteins are then degraded in the vacuole.