ER degradation of a misfolded luminal protein by the cytosolic ubiquitin-proteasome pathway

Science

Volumn 273, Issue 5282, 1996, Pages 1725-1728

  Hiller, Mark M ^a   Finger, Andreas ^a   Schweiger, Markus ^a   Wolf, Dieter H ^a  

^a UNIVERSITY OF STUTTGART   (Germany)

Author keywords

[No Author keywords available]

Indexed keywords

CARBOXYPEPTIDASE; PROTEASOME; UBIQUITIN;

ARTICLE; CELL ORGANELLE; ENDOPLASMIC RETICULUM; GENE MUTATION; GLYCOSYLATION; NONHUMAN; PRIORITY JOURNAL; PROTEIN CONFORMATION; PROTEIN DEGRADATION; PROTEIN FOLDING; PROTEIN SECRETION;

EID: 0029838640     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.273.5282.1725     Document Type: Article

References (53)

1. N. K. Pryer, L. J. Wuestehube, R. Schekman, Annu. Rev. Biochem. 61, 471 (1992); J. R. Gaut and L. M. Hendershot. Curr. Opin. Cell Biol. 5, 589 (1993).
2. N. K. Pryer, L. J. Wuestehube, R. Schekman, Annu. Rev. Biochem. 61, 471 (1992); J. R. Gaut and L. M. Hendershot. Curr. Opin. Cell Biol. 5, 589 (1993).
3. J. E. Rothman, Cell 59, 591 (1989); M. J. Gething and J. Sambrook, Nature 355, 33 (1992); R. W. Doms, R. A. Lamb, J. K. Rose, A. Helenius, Virology 193, 545 (1993); F. U. Hartl, R. Hlodan, T. Langer, Trends Biochem. Sci. 19, 20 (1994).
4. J. E. Rothman, Cell 59, 591 (1989); M. J. Gething and J. Sambrook, Nature 355, 33 (1992); R. W. Doms, R. A. Lamb, J. K. Rose, A. Helenius, Virology 193, 545 (1993); F. U. Hartl, R. Hlodan, T. Langer, Trends Biochem. Sci. 19, 20 (1994).
5. J. E. Rothman, Cell 59, 591 (1989); M. J. Gething and J. Sambrook, Nature 355, 33 (1992); R. W. Doms, R. A. Lamb, J. K. Rose, A. Helenius, Virology 193, 545 (1993); F. U. Hartl, R. Hlodan, T. Langer, Trends Biochem. Sci. 19, 20 (1994).
6. J. E. Rothman, Cell 59, 591 (1989); M. J. Gething and J. Sambrook, Nature 355, 33 (1992); R. W. Doms, R. A. Lamb, J. K. Rose, A. Helenius, Virology 193, 545 (1993); F. U. Hartl, R. Hlodan, T. Langer, Trends Biochem. Sci. 19, 20 (1994).
7. J. S. Bonifacino and R. D. Klausner, in Cellular Proteolytic Systems, A. Ciechanover and A. L. Schwartz, Eds. (Wiley-Liss, New York, 1994), vol. 15, pp. 137-160.
8. S. J. Lippincott, J. S. Bonifacino, L. C. Yuan, R. D. Klausner, Cell 54, 209 (1988); R. D. Klausner and R. Sitia, ibid. 62, 611 (1990); J. S. Bonifacino and S. J. Lippincott, Curr. Opin. Cell Biol. 3, 592 (1991).
9. S. J. Lippincott, J. S. Bonifacino, L. C. Yuan, R. D. Klausner, Cell 54, 209 (1988); R. D. Klausner and R. Sitia, ibid. 62, 611 (1990); J. S. Bonifacino and S. J. Lippincott, Curr. Opin. Cell Biol. 3, 592 (1991).
10. S. J. Lippincott, J. S. Bonifacino, L. C. Yuan, R. D. Klausner, Cell 54, 209 (1988); R. D. Klausner and R. Sitia, ibid. 62, 611 (1990); J. S. Bonifacino and S. J. Lippincott, Curr. Opin. Cell Biol. 3, 592 (1991).
11. C. L. Ward, S. Omura, R. R. Kopito, Cell 83, 121 (1995); T. J. Jensen et al., ibid., p. 129.
12. C. L. Ward, S. Omura, R. R. Kopito, Cell 83, 121 (1995); T. J. Jensen et al., ibid., p. 129.
13. A. Finger, M. Knop, D. H. Wolf, Eur. J Biochem. 218, 565 (1993).
14. M. Knop, A. Finger, T. Braun, K. Hellmuth, D. H. Wolf, EMBO J. 15, 753 (1996).
15. M. M. Hiller, unpublished results.
16. The complementing plasmid, containing a 3.95-kb insert, was isolated by transforming the der2-1 mutant strain (31) with a genomic library contained in plasmid yCPlac111 [F. Cvrckova and K. Nasmyth, EMBO J. 12, 5277 (1993)] and screening for complementing colonies as described (7). Sequencing this insert from both ends and subsequent search in the EMBL database revealed the presence of the genes MSS1, UBC7 (QR18), and MAC1. Only the 1.1-kb Eco RI-Pst I subclone containing UBC7 (10, 11) was able to complement the der2-1 mutation. The presence of UBC7 was verified by sequencing. A cross of the der2-1 mutant with a ubc7 deletion strain (W303-CQ) (31) did not lead to complementation of the mutant phenotype in the resulting diploid. Sporulation of this diploid produced four spores, all of which exhibited the mutant phenotype, indicating that DER2 is identical to UBC7 (QR18) (8).
17. A. Vassal, A. Boulet, E. Decoster, G. Faye, Biochim. Biophys. Acta 1132, 211 (1992).
18. J. Jungmann, H. A. Reins, C. Schobert, S. Jentsch, Nature 361, 369 (1993).
19. P. Chen, P. Johnson, T. Sommer, S. Jentsch, M. Hochstrasser, Cell 74, 357 (1993).
20. T. Sommer and S. Jentsch, Nature 365, 176 (1993).
21. W. Seufert and S. Jentsch, EMBO J. 9, 543 (1990).
22. V. Chau et al., Science 243, 1576 (1989).
23. T. Arnason and M. J. Ellison, Mol. Cell. Biol. 14, 7876 (1994).
24. M. Hochstrasser, M. J. Ellison, V. Chau, A. Varshavsky, Proc. Natl. Acad. Sci. U.S.A. 88, 4606 (1991).
25. S. M. Schork, M. Thumm, D. H. Wolf, J. Biol. Chem. 270, 26446 (1995).
26. M. Hochstrasser, Curr. Opin. Cell Biol. 7, 215 (1995); W. Hilt and D. H. Wolf, Trends Biochem. Sci. 21, 96 (1996).
27. M. Hochstrasser, Curr. Opin. Cell Biol. 7, 215 (1995); W. Hilt and D. H. Wolf, Trends Biochem. Sci. 21, 96 (1996).
28. W. Heinemeyer, J. A. Kleinschmidt, J. Saidowsky, C. Escher, D. H. Wolf, EMBO J. 10, 555 (1991); W Hilt, C. Enenkel, A. Gruhler, T. Singer, D. H. Wolf, J. Biol. Chem. 268, 3479 (1993).
29. W. Heinemeyer, J. A. Kleinschmidt, J. Saidowsky, C. Escher, D. H. Wolf, EMBO J. 10, 555 (1991); W Hilt, C. Enenkel, A. Gruhler, T. Singer, D. H. Wolf, J. Biol. Chem. 268, 3479 (1993).
30. W. Heinemeyer, A. Gruhler, V. Mohrle, Y Mahe, D. H. Wolf, J. Biol. Chem. 268, 5115 (1993).
31. M. Ghislain, A. Udvardy, C. Mann. Nature 366, 358 (1993).
32. D. M. Rubin et al., ibid. 379, 655 (1996).
33. A. J. Rivett and E. Knecht, Curr. Biol. 3, 127 (1993); A. Palmer, G. G. Mason, J. M. Paramio, E. Knecht, A. J. Rivett, Eur. J. Cell Biol. 64, 163 (1994); U. Vallon and U. Kull, Protoplasma 182, 15 (1994).
34. A. J. Rivett and E. Knecht, Curr. Biol. 3, 127 (1993); A. Palmer, G. G. Mason, J. M. Paramio, E. Knecht, A. J. Rivett, Eur. J. Cell Biol. 64, 163 (1994); U. Vallon and U. Kull, Protoplasma 182, 15 (1994).
35. A. J. Rivett and E. Knecht, Curr. Biol. 3, 127 (1993); A. Palmer, G. G. Mason, J. M. Paramio, E. Knecht, A. J. Rivett, Eur. J. Cell Biol. 64, 163 (1994); U. Vallon and U. Kull, Protoplasma 182, 15 (1994).
36. A. L. Schwartz, A. Ciechanover, R. A. Brandt, H. J. Geuze, EMBO J. 7, 2961 (1988).
37. K. Römisch and R. Schekman, Proc. Natl. Acad. Sci. U.S.A. 89, 7227 (1992); J. Roelse, M. Gromme, F. Momburg, G. Hammerling, J. Neetjes, J. Exp. Med. 180, 1591 (1994).
38. K. Römisch and R. Schekman, Proc. Natl. Acad. Sci. U.S.A. 89, 7227 (1992); J. Roelse, M. Gromme, F. Momburg, G. Hammerling, J. Neetjes, J. Exp. Med. 180, 1591 (1994).
39. M. D. Rose, L. M. Misra, J. P. Vogel, Cell 57, 1211 (1989).
40. A. A. McCracken and J. L. Brodsky, J. Cell Biol. 132, 291 (1996).
41. E. J. H. J. Wiertz et al., Cell 84, 769 (1996).
42. 35S]methionine. Growth, labeling, and chase conditions, as well as all other experimental procedures (cell breakage, immunoprecipitation, and SDS-PAGE), were as described (6).
43. + 12 contain WT-Ub, Ub-R48 and HA-Ub under the control of the CUP1 promoter on a 2μ plasmid (17). The plasmids containing Ub-R29 and Ub-R63 are derivatives of Yep96 (16).
44. + 12 contain WT-Ub, Ub-R48 and HA-Ub under the control of the CUP1 promoter on a 2μ plasmid (17). The plasmids containing Ub-R29 and Ub-R63 are derivatives of Yep96 (16).
45. + 12 contain WT-Ub, Ub-R48 and HA-Ub under the control of the CUP1 promoter on a 2μ plasmid (17). The plasmids containing Ub-R29 and Ub-R63 are derivatives of Yep96 (16).
46. + 12 contain WT-Ub, Ub-R48 and HA-Ub under the control of the CUP1 promoter on a 2μ plasmid (17). The plasmids containing Ub-R29 and Ub-R63 are derivatives of Yep96 (16).
47. + 12 contain WT-Ub, Ub-R48 and HA-Ub under the control of the CUP1 promoter on a 2μ plasmid (17). The plasmids containing Ub-R29 and Ub-R63 are derivatives of Yep96 (16).
48. + 12 contain WT-Ub, Ub-R48 and HA-Ub under the control of the CUP1 promoter on a 2μ plasmid (17). The plasmids containing Ub-R29 and Ub-R63 are derivatives of Yep96 (16).
49. + 12 contain WT-Ub, Ub-R48 and HA-Ub under the control of the CUP1 promoter on a 2μ plasmid (17). The plasmids containing Ub-R29 and Ub-R63 are derivatives of Yep96 (16).
50. 8 cells) was separated on an 8% SDS-polyacrylamide gel and transferred to nitrocellulose membranes according to standard methods. HA-Ub was detected with the 12CA5 antibody to HA and enhanced chemiluminescence (ECL; Amersham). Kar2p was similarly detected with specific polyclonal antibodies.
51. 600 units of cells. From the lime of lysis, all material was maintained on ice. Membranes were separated from the soluble fraction by centrifugation at 20,000g for 30 min at 4°C. For protease treatment, the pellet was resuspended and trypsin was added to a concentration of 0.5 mg/ml. The sample was incubated for 30 min on ice (if added, Triton X-100 was present at 1% during this incubation). All treatments were terminated by precipitation of proteins with 10% TCA for 10 min on ice Solubilization of the TCA pellet, immunoprecipitation of CPY* antigenic material, SDS-PAGE, and defection of HA-Ub-conjugated CPY* were performed as described (32).
52. 600 unit of cells) for 3 hours at room temperature with gentle agitation. The Con A-Sepharose was separated by centrifugation (1500g, 3 min) and washed twice with Con A buffer. Proteins of the combined supernatant and washes were precipitated with 10% TCA for 10 min on ice and solubilized in UREA buffer (32). as were the proteins bound to Con A-Sepharose. SDS-PAGE and detection of HA-Ubconjugated CPY* were performed as described (32).
53. We thank H. L. Chiang, S. Fuller, W. Heinemeyer, W. Hilt, M. Knop, and C. Mann for providing strains; F. Cvrckova and K. Nasmyth for the CEN-LEU2 library; M. J. Ellison, G. Faye, M. Hochstrasser, and S. Jentsch for providing plasmids; and H. Rudolph and R. Schekman for the antibodies to HA and to Kar2p, respectively. Supported by the Bundesministerium für Forschung und Technologie (ZSP Stuttgart grant B 3.7 U) and the Fonds der Chemischen Industrie, Frankfurt.