Ultrastructural analysis of the autophagic process in yeast: detection of autophagosomes and their characterization

doi:10.1083/jcb.124.6.903

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Ultrastructural analysis of the autophagic process in yeast: detection of autophagosomes and their characterization

M Baba, K Takeshige, N Baba and Y Ohsumi
Department of Biology, College of Arts and Sciences, University of Tokyo, Japan.

Under nutrient-deficient conditions, the yeast S. cerevisiae sequesters its own cytoplasmic components into vacuoles in the form of "autophagic bodies" (Takeshige, K., M. Baba, S. Tsuboi, T. Noda, and Y. Ohsumi. 1992. J. Cell Biol. 119:301-311). Immunoelectron microscopy showed that two cytosolic marker enzymes, alcohol dehydrogenase and phosphoglycerate kinase, are present in the autophagic bodies at the same densities as in the cytosol, but are not present in vacuolar sap, suggesting that cytosolic enzymes are also taken up into the autophagic bodies. To understand this process, we performed morphological analyses by transmission and immunological electron microscopies using a freeze- substitution fixation method. Spherical structures completely enclosed in a double membrane were found near the vacuoles of protease-deficient mutant cells when the cells were shifted to nutrient-starvation media. Their size, membrane thickness, and contents of double membrane- structures corresponded well with those of autophagic bodies. Sometimes these double membrane structures were found to be in contact with the vacuolar membrane. Furthermore their outer membrane was occasionally seen to be continuous with the vacuolar membrane. Histochemical staining of carbohydrate strongly suggested that the structures with double membranes fused with the vacuoles. These results indicated that these structures are precursors of autophagic bodies, "autophagosomes" in yeast. All the data obtained suggested that the autophagic process in yeast is essentially similar to that of the lysosomal system in mammalian cells.
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Lang, T., Reiche, S., Straub, M., Bredschneider, M., Thumm, M. (2000). Autophagy and the cvt Pathway Both Depend on AUT9. J. Bacteriol. 182: 2125-2133 [Abstract] [Full Text]
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Huang, W.-P., Scott, S. V., Kim, J., Klionsky, D. J. (2000). The Itinerary of a Vesicle Component, Aut7p/Cvt5p, Terminates in the Yeast Vacuole via the Autophagy/Cvt Pathways. J. Biol. Chem. 275: 5845-5851 [Abstract] [Full Text]
Noda, T., Kim, J., Huang, W.-P., Baba, M., Tokunaga, C., Ohsumi, Y., Klionsky, D. J. (2000). Apg9p/Cvt7p Is an Integral Membrane Protein Required for Transport Vesicle Formation in the Cvt and Autophagy Pathways. JCB 148: 465-480 [Abstract] [Full Text]
Suriapranata, I, Epple, U., Bernreuther, D, Bredschneider, M, Sovarasteanu, K, Thumm, M (2000). The breakdown of autophagic vesicles inside the vacuole depends on Aut4p. J. Cell Sci. 113: 4025-4033 [Abstract]
Kirisako, T., Baba, M., Ishihara, N., Miyazawa, K., Ohsumi, M., Yoshimori, T., Noda, T., Ohsumi, Y. (1999). Formation Process of Autophagosome Is Traced with Apg8/Aut7p in Yeast. JCB 147: 435-446 [Abstract] [Full Text]
Ueno, T., Ishidoh, K., Mineki, R., Tanida, I., Murayama, K., Kadowaki, M., Kominami, E. (1999). Autolysosomal Membrane-associated Betaine Homocysteine Methyltransferase. LIMITED DEGRADATION FRAGMENT OF A SEQUESTERED CYTOSOLIC ENZYME MONITORING AUTOPHAGY. J. Biol. Chem. 274: 15222-15229 [Abstract] [Full Text]
Kim, J., Dalton, V. M., Eggerton, K. P., Scott, S. V., Klionsky, D. J. (1999). Apg7p/Cvt2p Is Required for the Cytoplasm-to-Vacuole Targeting, Macroautophagy, and Peroxisome Degradation Pathways. Mol. Biol. Cell 10: 1337-1351 [Abstract] [Full Text]
Yuan, W., Strømhaug, P. E., Dunn, W. A. Jr. (1999). Glucose-induced Autophagy of Peroxisomes in Pichia pastoris Requires a Unique E1-like Protein. Mol. Biol. Cell 10: 1353-1366 [Abstract] [Full Text]
Tanida, I., Mizushima, N., Kiyooka, M., Ohsumi, M., Ueno, T., Ohsumi, Y., Kominami, E. (1999). Apg7p/Cvt2p: A Novel Protein-activating Enzyme Essential for Autophagy. Mol. Biol. Cell 10: 1367-1379 [Abstract] [Full Text]
Hutchins, M., Veenhuis, M, Klionsky, D. (1999). Peroxisome degradation in Saccharomyces cerevisiae is dependent on machinery of macroautophagy and the Cvt pathway. J. Cell Sci. 112: 4079-4087 [Abstract]
Biederbick, A, Rose, S, Elsasser, H. (1999). A human intracellular apyrase-like protein, LALP70, localizes to lysosomal/autophagic vacuoles. J. Cell Sci. 112: 2473-2484 [Abstract]
Schaible, U., Schlesinger, P., Steinberg, T., Mangel, W., Kobayashi, T, Russell, D. (1999). Parasitophorous vacuoles of Leishmania mexicana acquire macromolecules from the host cell cytosol via two independent routes. J. Cell Sci. 112: 681-693 [Abstract]
Mizushima, N., Sugita, H., Yoshimori, T., Ohsumi, Y. (1998). A New Protein Conjugation System in Human. THE COUNTERPART OF THE YEAST Apg12p CONJUGATION SYSTEM ESSENTIAL FOR AUTOPHAGY. J. Biol. Chem. 273: 33889-33892 [Abstract] [Full Text]
Kametaka, S., Okano, T., Ohsumi, M., Ohsumi, Y. (1998). Apg14p and Apg6/Vps30p Form a Protein Complex Essential for Autophagy in the Yeast, Saccharomyces cerevisiae. J. Biol. Chem. 273: 22284-22291 [Abstract] [Full Text]
Brouquisse, R., Gaudillère, J.-P., Raymond, P. (1998). Induction of a Carbon-Starvation-Related Proteolysis in Whole Maize Plants Submitted to Light/Dark Cycles and to Extended Darkness. Plant Physiol. 117: 1281-1291 [Abstract] [Full Text]
Sakai, Y., Koller, A., Rangell, L. K., Keller, G. A., Subramani, S. (1998). Peroxisome Degradation by Microautophagy in Pichia pastoris: Identification of Specific Steps and Morphological Intermediates. JCB 141: 625-636 [Abstract] [Full Text]
Klionsky, D. J. (1998). Nonclassical Protein Sorting to the Yeast Vacuole. J. Biol. Chem. 273: 10807-10810 [Full Text]
Bryant, N. J., Stevens, T. H. (1998). Vacuole Biogenesis in Saccharomyces cerevisiae: Protein Transport Pathways to the Yeast Vacuole. Microbiol. Mol. Biol. Rev. 62: 230-247 [Abstract] [Full Text]
Noda, T., Ohsumi, Y. (1998). Tor, a Phosphatidylinositol Kinase Homologue, Controls Autophagy in Yeast. J. Biol. Chem. 273: 3963-3966 [Abstract] [Full Text]
Campbell, C., Thorsness, P. (1998). Escape of mitochondrial DNA to the nucleus in yme1 yeast is mediated by vacuolar-dependent turnover of abnormal mitochondrial compartments. J. Cell Sci. 111: 2455-2464 [Abstract]
SUBRAMANI, S. (1998). Components Involved in Peroxisome Import, Biogenesis, Proliferation, Turnover, and Movement. Physiol. Rev. 78: 171-188 [Abstract] [Full Text]
Baba, M., Osumi, M., Scott, S. V., Klionsky, D. J., Ohsumi, Y. (1997). Two Distinct Pathways for Targeting Proteins from the Cytoplasm to the Vacuole/Lysosome. JCB 139: 1687-1695 [Abstract] [Full Text]
Rieder, S. E., Emr, S. D. (1997). A Novel RING Finger Protein Complex Essential for a Late Step in Protein Transport to the Yeast Vacuole. Mol. Biol. Cell 8: 2307-2327 [Abstract] [Full Text]
Darsow, T., Rieder, S. E., Emr, S. D. (1997). A Multispecificity Syntaxin Homologue, Vam3p, Essential for Autophagic and Biosynthetic Protein Transport to the Vacuole. JCB 138: 517-529 [Abstract] [Full Text]
Scott, S. V., Baba, M., Ohsumi, Y., Klionsky, D. J. (1997). Aminopeptidase I Is Targeted to the Vacuole by a Nonclassical Vesicular Mechanism. JCB 138: 37-44 [Abstract] [Full Text]
Kim, J., Scott, S. V., Oda, M. N., Klionsky, D. J. (1997). Transport of a Large Oligomeric Protein by the Cytoplasm to Vacuole Protein Targeting Pathway. JCB 137: 609-618 [Abstract] [Full Text]
Yuan, W, Tuttle, D., Shi, Y., Ralph, G., Dunn, W. (1997). Glucose-induced microautophagy in Pichia pastoris requires the alpha-subunit of phosphofructokinase. J. Cell Sci. 110: 1935-1945 [Abstract]
Harding, T. M., Hefner-Gravink, A., Thumm, M., Klionsky, D. J. (1996). Genetic and Phenotypic Overlap between Autophagy and the Cytoplasm to Vacuole Protein Targeting Pathway. J. Biol. Chem. 271: 17621-17624 [Abstract] [Full Text]
Tuttle, D., Dunn, W. (1995). Divergent modes of autophagy in the methylotrophic yeast Pichia pastoris. J. Cell Sci. 108: 25-35 [Abstract]
Tanida, I., Tanida-Miyake, E., Ueno, T., Kominami, E. (2001). The Human Homolog of Saccharomyces cerevisiae Apg7p Is a Protein-activating Enzyme for Multiple Substrates Including Human Apg12p, GATE-16, GABARAP, and MAP-LC3. J. Biol. Chem. 276: 1701-1706 [Abstract] [Full Text]
Komatsu, M., Tanida, I., Ueno, T., Ohsumi, M., Ohsumi, Y., Kominami, E. (2001). The C-terminal Region of an Apg7p/Cvt2p Is Required for Homodimerization and Is Essential for Its E1 Activity and E1-E2 Complex Formation. J. Biol. Chem. 276: 9846-9854 [Abstract] [Full Text]
Shintani, T., Suzuki, K., Kamada, Y., Noda, T., Ohsumi, Y. (2001). Apg2p Functions in Autophagosome Formation on the Perivacuolar Structure. J. Biol. Chem. 276: 30452-30460 [Abstract] [Full Text]
Toyooka, K., Okamoto, T., Minamikawa, T. (2001). Cotyledon cells of Vigna mungo seedlings use at least two distinct autophagic machineries for degradation of starch granules and cellular components. JCB 154: 973-982 [Abstract] [Full Text]