Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

A proteolytic system that compensates for loss of proteasome function

Nature volume 392, pages 618–622 (1998)Cite this article

Abstract

Proteolysis is essential for the execution of many cellular functions. These include removal of incorrectly folded or damaged proteins1, the activation of transcription factors2, the ordered degradation of proteins involved in cell cycle control3, and the generation of peptides destined for presentation by class I molecules of the major histocompatibility complex4. A multisubunit protease complex, the proteasome5, accomplishes these tasks. Here we show that in mammalian cells inactivation of the proteasome by covalent inhibitors allows the outgrowth of inhibitor-resistant cells. The growth of such adapted cells is apparently maintained by the induction of other proteolytic systems that compensate for the loss of proteasomal activity.

This is a preview of subscription content

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

Buy

All prices are NET prices.

Figure 1: Covalent modification of proteasomes in EL-4 cells cultured in the presence of NLVS blocks enzymatic activity.
Figure 2: Peptidase activity from EL-4-adapted and control fractions resolved by gel filtration (see Methods).
Figure 3: Synthesis and assembly of proteasome subunits is normal in adapted cells.
Figure 4: Adapted EL-4 cells generate ligands for stable assembly and intracellular transport of MHC class I molecules.
Figure 5: AAF-MCA hydrolysis is increased in adapted cells.
Figure 6: AAF-MCA hydrolysing activity elutes before the proteasome, indicating that it has a higher M r.

References

  1. Etlinger, J. D. & Goldberg, A. L. Asoluble ATP-dependent proteolytic system responsible for the degradation of abnormal proteins in reticulocytes. Proc. Natl Acad. Sci. USA 74, 54–58 (1977).

    ADS  CAS  Article  Google Scholar 

  2. Palombella, V. J., Rando, O. J., Goldberg, A. L. & Maniatis, T. The ubiquitin-proteasome pathway is required for processing the NF-κB1 precursor protein and the activation of NF-κB. Cell 78, 773–785 (1994).

    CAS  Article  Google Scholar 

  3. Glotzer, M., Murray, A. W. & Kirschner, M. W. Cyclin is degraded by the ubiquitin pathway. Nature 349, 132–138 (1991).

    ADS  CAS  Article  Google Scholar 

  4. Rock, K. L.et al. Inhibition of the proteasome block the degradation of most cell proteins and the generation of peptides presented on MHC class I molecules. Cell 78, 761–771 (1994).

    CAS  Article  Google Scholar 

  5. Coux, O., Tanaka, K. & Goldberg, A. Structure and functions of the 20S and 26S proteasomes. Annu. Rev. Biochem. 65, 801–847 (1996).

    CAS  Article  Google Scholar 

  6. Seemuller, E.et al. The proteasome from Thermoplasma acidophilum : A threonine protease. Science 268, 579–582 (1995).

    ADS  CAS  Article  Google Scholar 

  7. Löwe, J.et al. Crystal structure of the 20S proteasome from the Archaeon T. acidophilum at 3.4å resolution. Science 268, 533–539 (1995).

    ADS  Article  Google Scholar 

  8. Omura, S.et al. Lactacystin, a novel microbial metabolite, induces neuritogenesis of neuroblastoma cells. J. Antibiot. 44, 113–116 (1991).

    CAS  Article  Google Scholar 

  9. Fenteany, G.et al. Inhibition of proteasome activities and subunit-specific amino-terminal threonine modification by lactacystin. Science 268, 726–730 (1995).

    ADS  CAS  Article  Google Scholar 

  10. Dick, L. R.et al. Mechanistic studies on the inactivation of the proteasome by lactacystin. J. Biol. Chem. 271, 7273–7276 (1996).

    CAS  Article  Google Scholar 

  11. Groll, M.et al. Structure of 20S proteasome from yeast at 2.4 å resolution. Nature 386, 463–471 (1997).

    ADS  CAS  Article  Google Scholar 

  12. Bogyo, M.et al. Covalent modification of the active site Thr of proteasomeal β-subunits and the E. coli homolog HslV by a new class of inhibitors. Proc. Natl Acad. Sci.. USA 94, 6629–6634 (1997).

    ADS  CAS  Article  Google Scholar 

  13. Nandi, D., Woodward, E., Ginsburg, D. B. & Monaco, J. J. Intermediates in the formation of mouse 20S proteasomes: implications for the assembly of precursor beta subunits. EMBO J. 16, 5363–5375 (1997).

    CAS  Article  Google Scholar 

  14. Driscoll, J., Brown, M. G., Finley, D. & Monaco, J. J. MHC-linked LMP gene products specifically alter peptidase activities of the proteasome. Nature 365, 262–264 (1993).

    ADS  CAS  Article  Google Scholar 

  15. Gaczynska, M., Rock, K. L. & Goldberg, A. γ-Interferon and expression of MHC genes regulate peptide hydrolysis by proteasomes. Nature 365, 264–267 (1993).

    ADS  CAS  Article  Google Scholar 

  16. Craiu, A. C.et al. Lactacystin and clasto-lactacystin β-lactone modify multiple proteasome β subunits and inhibit intracellular protein degradation and MHC class I antigen presentation. J. Biol. Chem. 272, 13437–13445 (1997).

    CAS  Article  Google Scholar 

  17. Bai, A. & Forman, J. The effect of proteasome inhibitor lactacystin on the presentation of transporter associated with antigen processing (TAP)-dependent and TAP-independent peptide epitopes by class I molecules. J. Immunol. 159, 2139–2146 (1007).

    Google Scholar 

  18. Vinitsky, A.et al. The generation of MHC class I-associated peptides is only partially inhibited by proteasome inhibitors: involvement of non-proteasomal cytosolic proteases in antigen processing? J. Immunol. 159, 554–564 (1997).

    CAS  PubMed  Google Scholar 

  19. Hilt, W. & Wolf, D. H. Proteasomes of the yeast S. cerevisiae : genes, structure and function. Mol. Biol. Rep. 21, 3–10 (1995).

    CAS  Article  Google Scholar 

  20. Ghislain, M., Udvardy, A. & Mann, C.et al. S. cerevisiae 26S protease mutants arrest cell division in G2/metaphase. Nature 366, 358–362 (1993).

    ADS  CAS  Article  Google Scholar 

  21. Gordon, C., McGurk, G., Dilion, P., Rosen, C. & Hastie, N. D. Defective mitosis due to a mutation in the gene for a fission yeast 26S protease subunit. Nature 366, 355–357 (1993).

    ADS  CAS  Article  Google Scholar 

  22. Tamura, T.et al. Tricorn protease — the core of a modular proteolytic system. Science 274, 1385–1389 (1996).

    ADS  CAS  Article  Google Scholar 

  23. Kettner, C. & Shaw, E. Inactivation of trypsin-like enzymes with peptides of arginine chloromethyl ketone. Meth. Enzymol. 80, 826–842 (1981).

    CAS  Article  Google Scholar 

  24. Drexler, H. C. A. Activation of the cell death program by inhibition of proteasome function. Proc. Natl Acad. Sci. USA 94, 855–860 (1997).

    ADS  CAS  Article  Google Scholar 

  25. Hershko, A. & Ciechanover, A. The ubiquitin system for protein degradation. Annu. Rev. Biochem. 61, 761–807 (1992).

    CAS  Article  Google Scholar 

  26. King, R. W., Jackson, P. K. & Kirschner, M. W. Mitosis in transition. Cell 79, 563–571 (1994).

    CAS  Article  Google Scholar 

  27. Sheaff, R. & Roberts, J. M. End of the line: proteolytic degradation of cyclin-dependent kinase inhibitors. Chem. Biol. 3, 869–873 (1996).

    CAS  Article  Google Scholar 

  28. Hammerling, G. J., Rusch, E., Tada, N., Kimura, S. & Hammerling, U. Localization of allo-determinants on H-2b determined with monoclonal antibodies and H-2 mutant mice. Proc. Natl Acad. Sci. USA 79, 4737–4741 (1982).

    ADS  CAS  Article  Google Scholar 

  29. Ljunggren, H.-G.et al. Empty MHC class I molecules come out in the cold. Nature 346, 476–480 (1990).

    ADS  CAS  Article  Google Scholar 

Download references

Acknowledgements

This work was supported by grants from the NIH and Boehringer Ingelheim. R. G. is a fellow of the The Swedish Foundation for International Cooperation in Research and High Education (STINT) Stockholm.

Author information

Author notes

  1. Rickard Glas, Matthew Bogyo & Hidde L. Ploegh

    Present address: Department of Pathology, Harvard Medical School, 200 Longwood Avenue, Boston, Massachusetts, 02115, USA

  2. John S. McMaster

    Present address: University of Utah Department of Biochemistry, Salt Lake City, Utah, 84132, USA

  3. Maria Gaczynska

    Present address: Department of Molecular Medicine, Institute of Biotechnology, Univesity of Texas Health Science Center, San Antonio, Texas, 78245, USA

  4. Rickard Glas and Matthew Bogyo: These authors contributed equally to this work

Affiliations

  1. Department for Biology, Center for Cancer Research, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, 02139-4307, Massachusetts, USA

    Rickard Glas, Matthew Bogyo, John S. McMaster, Maria Gaczynska & Hidde L. Ploegh

Authors
  1. Rickard Glas
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Matthew Bogyo
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. John S. McMaster
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Maria Gaczynska
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Hidde L. Ploegh
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hidde L. Ploegh.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Glas, R., Bogyo, M., McMaster, J. et al. A proteolytic system that compensates for loss of proteasome function. Nature 392, 618–622 (1998). https://doi.org/10.1038/33443

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/33443

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing