1887

Abstract

The multivesicular body (MVB) sorting pathway is required for a number of biological processes, including downregulation of cell-surface proteins and protein sorting into the vacuolar lumen. The function of this pathway requires endosomal sorting complexes required for transport (ESCRT) composed of class E vacuolar protein sorting (Vps) proteins in , many of which are conserved in . Of these, / (homologous to ) and (similar to ) have been identified as suppressors of sterility in Δ (), although their functions have not been uncovered to date. In this report, these two genes are shown to be required for vacuolar sorting of carboxypeptidase Y (CPY) and an MVB marker, the ubiquitin–GFP–carboxypeptidase S (Ub–GFP–CPS) fusion protein, despite the lack of the ubiquitin E2 variant domain in Sst6p. Disruption mutants of a variety of other class E homologues also had defects in sorting of CPY and Ub–GFP–CPS. has a mammalian AMSH homologue, . Phenotypic analyses suggested that Sst2p is a class E Vps protein. Taken together, these results suggest that sorting into multivesicular bodies is dependent on class E Vps proteins, including Sst2p, in .

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2007-08-01
2019-10-22
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References

  1. Agromayor, M. & Martin-Serrano, J. ( 2006; ). Interaction of AMSH with ESCRT-III and deubiquitination of endosomal cargo. J Biol Chem 281, 23083–23091.[CrossRef]
    [Google Scholar]
  2. Alfa, C., Fantes, P., Hyams, J., McLeod, M. & Warbrick, E. ( 1993; ). Experiments with Fission Yeast: a Laboratory Course Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  3. Alwan, H. A. J. & van Leeuwen, J. E. M. ( 2007; ). UBPY-mediated EGFR deubiquitination promotes EGFR degradation. J Biol Chem 282, 1658–1669.[CrossRef]
    [Google Scholar]
  4. Amerik, A. Y., Nowak, J., Swaminathan, S. & Hochstrasser, M. ( 2000; ). The Doa4 deubiquitinating enzyme is functionally linked to the vacuolar protein-sorting and endocytic pathways. Mol Biol Cell 11, 3365–3380.[CrossRef]
    [Google Scholar]
  5. Babst, M. ( 2005; ). A protein's final ESCRT. Traffic 6, 2–9.[CrossRef]
    [Google Scholar]
  6. Babst, M., Sato, T. K., Banta, L. M. & Emr, S. D. ( 1997; ). Endosomal transport function in yeast requires a novel AAA-type ATPase, Vps4p. EMBO J 16, 1820–1831.[CrossRef]
    [Google Scholar]
  7. Babst, M., Wendland, B., Estepa, E. J. & Emr, S. D. ( 1998; ). The Vps4p AAA ATPase regulates membrane association of a Vps protein complex required for normal endosome function. EMBO J 17, 2982–2993.[CrossRef]
    [Google Scholar]
  8. Babst, M., Odorizzi, G., Estepa, E. J. & Emr, S. D. ( 2000; ). Mammalian tumor susceptibility gene 101 (TSG101) and the yeast homologue, Vps23p, both function in late endosomal trafficking. Traffic 1, 248–258.[CrossRef]
    [Google Scholar]
  9. Babst, M., Katzmann, D. J., Estepa-Sabal, E. J., Meerloo, T. & Emr, S. D. ( 2002; ). ESCRT-III: an endosome-associated heterooligomeric protein complex required for MVB sorting. Dev Cell 3, 271–282.[CrossRef]
    [Google Scholar]
  10. Bache, K. G., Slagsvold, T., Cabezas, A., Rosendal, K. R., Raiborg, C. & Stenmark, H. ( 2004; ). The growth-regulatory protein HCRP1/hVps37A is a subunit of mammalian ESCRT-I and mediates receptor down-regulation. Mol Biol Cell 15, 4337–4346.[CrossRef]
    [Google Scholar]
  11. Banta, L. M., Robinson, J. S., Klionsky, D. J. & Emr, S. D. ( 1988; ). Organelle assembly in yeast: characterization of yeast mutants defective in vacuolar biogenesis and protein sorting. J Cell Biol 107, 1369–1383.[CrossRef]
    [Google Scholar]
  12. Bilodeau, P. S., Urbanowski, J. L., Winistorfer, S. C. & Piper, R. C. ( 2002; ). The Vps27p-Hse1p complex binds ubiquitin and mediates endosomal protein sorting. Nat Cell Biol 4, 534–539.
    [Google Scholar]
  13. Bishop, N. & Woodman, P. ( 2001; ). TSG101/mammalian VPS23 and mammalian VPS28 interact directly and are recruited to VPS4-induced endosomes. J Biol Chem 276, 11735–11742.[CrossRef]
    [Google Scholar]
  14. Bowers, K. & Stevens, T. H. ( 2005; ). Protein transport from the late Golgi to the vacuole in the yeast Saccharomyces cerevisiae. Biochim Biophys Acta 1744, 438–454.[CrossRef]
    [Google Scholar]
  15. Bowers, K., Piper, S. C., Edeling, M. A., Gray, S. R., Owen, D. J., Lehner, P. J. & Luzio, J. P. ( 2006; ). Degradation of endocytosed epidermal growth factor and virally ubiquitinated major histocompatibility complex class I is independent of mammalian ESCRT II. J Biol Chem 281, 5094–5105.[CrossRef]
    [Google Scholar]
  16. Burd, C. G. & Emr, S. D. ( 1998; ). Phosphatidylinositol (3)-phosphate signaling mediated by specific binding to RING FYVE domains. Mol Cell 2, 157–162.[CrossRef]
    [Google Scholar]
  17. Cheng, H., Sugiura, R., Wu, W., Fujita, M., Lu, Y., Sio, S. O., Kawai, R., Takegawa, K., Shuntoh, H. & Kuno, T. ( 2002; ). Role of the Rab GTP-binding protein Ypt3 in the fission yeast exocytic pathway, and its connection to calcineurin function. Mol Biol Cell 13, 2963–2976.[CrossRef]
    [Google Scholar]
  18. Dupré, S. & Haguenauer-Tsapis, R. ( 2001; ). Deubiquitination in the endocytic pathway of yeast plasma membrane proteins: crucial role of Doa4p ubiquitin isopeptidase. Mol Cell Biol 21, 4482–4494.[CrossRef]
    [Google Scholar]
  19. Eastman, S. W., Martin-Serrano, J., Chung, W., Zang, T. & Bieniasz, P. D. ( 2005; ). Identification of human VPS37C, a component of endosomal sorting complex required for transport-I important for viral budding. J Biol Chem 280, 628–636.[CrossRef]
    [Google Scholar]
  20. Hicke, L. ( 1999; ). Gettin' down with ubiquitin: turning off cell-surface receptors, transporters and channels. Trends Cell Biol 9, 107–112.[CrossRef]
    [Google Scholar]
  21. Hirota, K., Tanaka, K., Watanabe, Y. & Yamamoto, M. ( 2001; ). Functional analysis of the C-terminal cytoplasmic region of the M-factor receptor in fission yeast. Genes Cells 6, 201–214.[CrossRef]
    [Google Scholar]
  22. Iwaki, T. & Takegawa, K. ( 2004; ). A set of loxP marker cassettes for Cre-mediated multiple gene disruption in Schizosaccharomyces pombe. Biosci Biotechnol Biochem 68, 545–550.[CrossRef]
    [Google Scholar]
  23. Iwaki, T., Tanaka, N., Takagi, H., Giga-Hama, Y. & Takegawa, K. ( 2004; ). Characterization of end4+ , a gene required for endocytosis in Schizosaccharomyces pombe. Yeast 21, 867–881.[CrossRef]
    [Google Scholar]
  24. Jin, Y., Mancuso, J. J., Cronembold, D. & Cande, W. Z. ( 2005; ). The fission yeast homolog of the human transcription factor EAP30 blocks meiotic spindle pole body amplification. Dev Cell 9, 63–73.[CrossRef]
    [Google Scholar]
  25. Katz, M., Shtiegman, K., Tal-Or, P., Yakir, L., Mosesson, Y., Harari, D., Machluf, Y., Asao, H., Jovin, T. & other authors ( 2002; ). Ligand-independent degradation of epidermal growth factor receptor involves receptor ubiquitylation and Hgs, an adaptor whose ubiquitin-interacting motif targets ubiquitylation by Nedd4. Traffic 3, 740–751.[CrossRef]
    [Google Scholar]
  26. Katzmann, D. J., Babst, M. & Emr, S. D. ( 2001; ). Ubiquitin-dependent sorting into the multivesicular body pathway requires the function of a conserved endosomal protein sorting complex, ESCRT-I. Cell 106, 145–155.[CrossRef]
    [Google Scholar]
  27. Katzmann, D. J., Sarkar, S., Chu, T., Audhya, A. & Emr, S. D. ( 2004; ). Multivesicular body sorting: ubiquitin ligase Rsp5 is required for the modification and sorting of carboxypeptidase S. Mol Biol Cell 15, 468–480.
    [Google Scholar]
  28. Koga, T., Onishi, M., Nakamura, Y., Hirata, A., Nakamura, T., Shimoda, C., Iwaki, T., Takegawa, K. & Fukui, Y. ( 2004; ). Sorting nexin homologues are targets of phosphatidylinositol 3-phosphate in sporulation of Schizosaccharomyces pombe. Genes Cells 9, 561–574.[CrossRef]
    [Google Scholar]
  29. Krsmanović, T., Pawelec, A., Sydor, T. & Kölling, R. ( 2005; ). Control of Ste6 recycling by ubiquitination in the early endocytic pathway in yeast. Mol Biol Cell 16, 2809–2821.[CrossRef]
    [Google Scholar]
  30. Lohi, O., Poussu, A., Mao, Y., Quiocho, F. & Lehto, V.-P. ( 2002; ). VHS domain – a longshoreman of vesicle lines. FEBS Lett 513, 19–23.[CrossRef]
    [Google Scholar]
  31. McCullough, J., Clague, M. J. & Urbé, S. ( 2004; ). AMSH is an endosome-associated ubiquitin isopeptidase. J Cell Biol 166, 487–492.[CrossRef]
    [Google Scholar]
  32. McEwen, R. K., Dove, S. K., Cooke, F. T., Painter, G. F., Holmes, A. B., Shisheva, A., Ohya, Y., Parker, P. J. & Michell, R. H. ( 1999; ). Complementation analysis in PrdInsP kinase-deficient yeast mutants demonstrates that Schizosaccharomyces pombe and murine Fab1p homologues are phosphatidylinositol 3-phosphate 5-kinases. J Biol Chem 274, 33905–33912.[CrossRef]
    [Google Scholar]
  33. Mitra, P., Zhang, Y., Rameh, L. E., Ivshina, M. P., McCollum, D., Nunnari, J. J., Hendricks, G. M., Kerr, M. L., Field, S. J. & other authors ( 2004; ). A novel phosphatidylinositol(3,4,5)P3 pathway in fission yeast. J Cell Biol 166, 205–211.[CrossRef]
    [Google Scholar]
  34. Mizuno, E., Iura, T., Mukai, A., Yoshimori, T., Kitamura, N. & Komada, M. ( 2005; ). Regulation of epidermal growth factor receptor down-regulation by UBPY-mediated deubiquitination at endosomes. Mol Biol Cell 16, 5163–5174.[CrossRef]
    [Google Scholar]
  35. Mizuno, E., Kobayashi, K., Yamamoto, A., Kitamura, N. & Komada, M. ( 2006; ). A deubiquitinating enzyme UBPY regulates the level of protein ubiquitination on endosomes. Traffic 7, 1017–1031.[CrossRef]
    [Google Scholar]
  36. Moreno, S., Klar, A. & Nurse, P. ( 1991; ). Molecular genetic analysis of fission yeast Schizosaccharomyces pombe. Methods Enzymol 194, 795–823.
    [Google Scholar]
  37. Morishita, M., Morimoto, F., Kitamura, K., Koga, T., Fukui, Y., Maekawa, H., Yamashita, I. & Shimoda, C. ( 2002; ). Phosphatidylinositol 3-phosphate 5-kinase is required for the cellular response to nutritional starvation and mating pheromone signals in Schizosaccharomyces pombe. Genes Cells 7, 199–215.[CrossRef]
    [Google Scholar]
  38. Morita, T. & Takegawa, K. ( 2004; ). A simple and efficient procedure for transformation of Schizosaccharomyces pombe. Yeast 21, 613–617.[CrossRef]
    [Google Scholar]
  39. Murray, J. M. & Johnson, D. I. ( 2001; ). The Cdc42p GTPase and its regulators of Nrf1p and Scd1p are involved in endocytic trafficking in the fission yeast Schizosaccharomyces pombe. J Biol Chem 276, 3004–3009.[CrossRef]
    [Google Scholar]
  40. Nakamura, T., Nakamura-Kubo, M., Hirata, A. & Shimoda, C. ( 2001; ). The Schizosaccharomyces pombe spo3 + gene is required for assembly of the forespore membrane and genetically interacts with psy1 +-encoding syntaxin-like protein. Mol Biol Cell 12, 3955–3972.[CrossRef]
    [Google Scholar]
  41. Odorizzi, G., Babst, M. & Emr, S. D. ( 1998; ). Fab1p PtdIns(3)P 5-kinase function essential for protein sorting in the multivesicular body. Cell 95, 847–858.[CrossRef]
    [Google Scholar]
  42. Okazaki, K., Okazaki, N., Kume, K., Jinno, S., Tanaka, K. & Okayama, H. ( 1990; ). High-frequency transformation method and library transducing vectors for cloning mammalian cDNAs by trans-complementation of Schizosaccharomyces pombe. Nucleic Acids Res 18, 6485–6489.[CrossRef]
    [Google Scholar]
  43. Onishi, M., Nakamura, Y., Koga, T., Takegawa, K. & Fukui, Y. ( 2003; ). Isolation of suppressor mutants of phosphatidylinositol 3-phosphate 5-kinase deficient cells in Schizosaccharomyces pombe. Biosci Biotechnol Biochem 67, 1772–1779.[CrossRef]
    [Google Scholar]
  44. Piper, R. C. & Luzio, J. P. ( 2001; ). Late endosomes: sorting and partitioning in multivesicular bodies. Traffic 2, 612–621.[CrossRef]
    [Google Scholar]
  45. Piper, R. C., Cooper, A. A., Yang, H. & Stevens, T. H. ( 1995; ). VPS27 controls vacuolar and endocytic traffic through a prevacuolar compartment in Saccharomyces cerevisiae. J Cell Biol 131, 603–617.[CrossRef]
    [Google Scholar]
  46. Polo, S., Sigismund, S., Faretta, M., Guidi, M., Capua, M. R., Bossi, G., Chen, H., De Camilli, P. & Di Fiore, P. P. ( 2002; ). A single motif responsible for ubiquitin recognition and monoubiquitination in endocytic proteins. Nature 416, 451–455.[CrossRef]
    [Google Scholar]
  47. Raymond, C. K., Howald-Stevenson, I., Vater, C. A. & Stevens, T. H. ( 1992; ). Morphological classification of the yeast vacuolar protein sorting mutants: evidence for a prevacuolar compartment in class E vps mutants. Mol Biol Cell 3, 1389–1402.[CrossRef]
    [Google Scholar]
  48. Reggiori, F. & Pelham, H. R. B. ( 2001; ). Sorting of proteins into multivesicular bodies: ubiquitin-dependent and -independent targeting. EMBO J 20, 5176–5186.[CrossRef]
    [Google Scholar]
  49. Rieder, S. E., Banta, L. M., Kohrer, K., McCaffery, J. M. & Emr, S. D. ( 1996; ). Multilamellar endosome-like compartment accumulates in the yeast vps28 vacuolar protein sorting mutant. Mol Biol Cell 7, 985–999.[CrossRef]
    [Google Scholar]
  50. Row, P. E., Prior, I. A., McCullough, J., Clague, M. J. & Urbé, S. ( 2006; ). The ubiquitin isopeptidase UBPY regulates endosomal ubiquitin dynamics and is essential for receptor down-regulation. J Biol Chem 281, 12618–12624.[CrossRef]
    [Google Scholar]
  51. Shih, S. C., Katzmann, D. J., Schnell, J. D., Sutanto, M., Emr, S. D. & Hicke, L. ( 2002; ). Epsins and Vps27p/Hrs contain ubiquitin-binding domains that function in receptor endocytosis. Nat Cell Biol 4, 389–393.[CrossRef]
    [Google Scholar]
  52. Soetens, O., De Craene, J.-O. & André, B. ( 2001; ). Ubiquitin is required for sorting to the vacuole of the yeast general amino acid permease, Gap1. J Biol Chem 276, 43949–43957.[CrossRef]
    [Google Scholar]
  53. Suga, M. & Hatakeyama, T. ( 2001; ). High efficiency transformation of Schizosaccharomyces pombe pretreated with thiol compounds by electroporation. Yeast 18, 1015–1021.[CrossRef]
    [Google Scholar]
  54. Suga, M., Isobe, M. & Hatakeyama, T. ( 2000; ). Cryopreservation of competent intact yeast cells for efficient electroporation. Yeast 16, 889–896.[CrossRef]
    [Google Scholar]
  55. Tabuchi, M., Iwaihara, O., Ohtani, Y., Ohuchi, N., Sakurai, J., Morita, T., Iwahara, S. & Takegawa, K. ( 1997; ). Vacuolar protein sorting in fission yeast: cloning, biosynthesis, transport, and processing of carboxypeptidase Y from Schizosaccharomyces pombe. J Bacteriol 179, 4179–4189.
    [Google Scholar]
  56. Takegawa, K., DeWald, D. B. & Emr, S. D. ( 1995; ). Schizosaccharomyces pombe Vps34p, a phosphatidylinositol-specific PI 3-kinase essential for normal cell growth and vacuole morphology. J Cell Sci 108, 3745–3756.
    [Google Scholar]
  57. Takegawa, K., Iwaki, T., Fujita, Y., Morita, T., Hosomi, A. & Tanaka, N. ( 2003; ). Vesicle-mediated protein transport pathways to the vacuole in Schizosaccharomyces pombe. Cell Struct Funct 28, 399–417.[CrossRef]
    [Google Scholar]
  58. Tanaka, N., Kaneko, K., Asao, H., Kasai, H., Endo, Y., Fujita, T., Takeshita, T. & Sugamura, K. ( 1999; ). Possible involvement of a novel STAM-associated molecule ‘AMSH’ in intracellular signal transduction mediated by cytokines. J Biol Chem 274, 19129–19135.[CrossRef]
    [Google Scholar]
  59. Tsang, H. T. H., Connell, J. W., Brown, S. E., Thompson, A., Reid, E. & Sanderson, C. M. ( 2006; ). A systematic analysis of human CHMP protein interactions: additional MIT domain-containing proteins bind to multiple components of the human ESCRT III complex. Genomics 88, 333–346.[CrossRef]
    [Google Scholar]
  60. Vida, T. A. & Emr, S. D. ( 1995; ). A new vital stain for visualizing vacuolar membrane dynamics and endocytosis in yeast. J Cell Biol 128, 779–792.[CrossRef]
    [Google Scholar]
  61. Winter, V. & Hauser, M.-T. ( 2006; ). Exploring the ESCRTing machinery in eukaryotes. Trends Plant Sci 11, 115–123.[CrossRef]
    [Google Scholar]
  62. Young, P., Deveraux, Q., Beal, R. E., Pickart, C. M. & Rechsteiner, M. ( 1998; ). Characterization of two polyubiquitin binding sites in the 26 S protease subunit 5a. J Biol Chem 273, 5461–5467.[CrossRef]
    [Google Scholar]
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