Abstract
The 26S proteasome is responsible for most intracellular proteolysis in eukaryotes1,2. Efficient substrate recognition relies on conjugation of substrates with multiple ubiquitin molecules and recognition of the polyubiquitin moiety by the 19S regulatory complex—a multisubunit assembly that is bound to either end of the cylindrical 20S proteasome core. Only unfolded proteins can pass through narrow axial channels into the central proteolytic chamber of the 20S core, so the attached polyubiquitin chain must be released to allow full translocation of the substrate polypeptide. Whereas unfolding is rate-limiting for the degradation of some substrates and appears to involve chaperone-like activities associated with the proteasome3,4,5, the importance and mechanism of degradation-associated deubiquitination has remained unclear. Here we report that the POH1 (also known as Rpn11 in yeast) subunit of the 19S complex is responsible for substrate deubiquitination during proteasomal degradation. The inability to remove ubiquitin can be rate-limiting for degradation in vitro and is lethal to yeast. Unlike all other known deubiquitinating enzymes (DUBs) that are cysteine proteases6,7, POH1 appears to be a Zn2+-dependent protease.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Hershko, A. & Ciechanover, A. The ubiquitin system. Annu. Rev. Biochem. 67, 425–479 (1998)
Zwickl, P., Baumeister, W. & Steven, A. Dis-assembly lines: the proteasome and related ATPase-assisted proteases. Curr. Opin. Struct. Biol. 10, 242–250 (2000)
Thrower, J. S., Hoffman, L., Rechsteiner, M. & Pickart, C. M. Recognition of the polyubiquitin proteolytic signal. EMBO J. 19, 94–102 (2000)
Braun, B. C. et al. The base of the proteasome regulatory particle exhibits chaperone-like activity. Nature Cell Biol. 1, 221–226 (1999)
Liu, C. et al. Conformational remodeling of proteasomal substrates by PA700, the 19S regulatory complex of the 26S proteasome. J. Biol. Chem. 277, 26815–26820 (2002)
D'Andrea, A. & Pellman, D. Deubiquitinating enzymes: a new class of biological regulators. Crit. Rev. Biochem. Mol. Biol. 33, 337–352 (1998)
Amerik, A. Y., Li, S. J. & Hochstrasser, M. Analysis of the deubiquitinating enzymes of the yeast Saccharomyces cerevisiae. Biol. Chem. 381, 981–992 (2000)
DeKoster, G. T. & Robertson, A. D. Thermodynamics of unfolding for Kazal-type serine protease inhibitors: entropic stabilization of ovomucoid first domain by glycosylation. Biochemistry 36, 2323–2331 (1997)
Johnson, E. S., Ma, P. C., Ota, I. M. & Varshavsky, A. A proteolytic pathway that recognizes ubiquitin as a degradation signal. J. Biol. Chem. 270, 17442–17456 (1995)
Hershko, A. & Rose, I. A. Ubiquitin-aldehyde: a general inhibitor of ubiquitin-recycling processes. Proc. Natl Acad. Sci. USA 84, 1829–1833 (1987)
Lam, Y. A., Xu, W., DeMartino, G. N. & Cohen, R. E. Editing of ubiquitin conjugates by an isopeptidase in the 26S proteasome. Nature 385, 737–740 (1997)
Wintrode, P. L., Makhatadze, G. I. & Privalov, P. L. Thermodynamics of ubiquitin unfolding. Proteins 18, 246–253 (1994)
Borodovsky, A. et al. A novel active site-directed probe specific for deubiquitylating enzymes reveals proteasome association of USP14. EMBO J. 20, 5187–5196 (2001)
Lam, Y. A., Lawson, T. G., Velayutham, M., Zweier, J. L. & Pickart, C. M. A proteasomal ATPase subunit recognizes the polyubiquitin degradation signal. Nature 416, 763–767 (2002)
Rubin, D. M., Glickman, M. H., Larsen, C. N., Dhruvakumar, S. & Finley, D. Active site mutants in the six regulatory particle ATPases reveal multiple roles for ATP in the proteasome. EMBO J. 17, 4909–4919 (1998)
Leggett, D. S. et al. Multiple associated proteins regulate proteasome structure and function. Mol. Cell (in the press)
Papa, F. R. & Hochstrasser, M. The yeast DOA4 gene encodes a deubiquitinating enzyme related to a product of the human tre-2 oncogene. Nature 366, 313–319 (1993)
Papa, F. R., Amerik, A. Y. & Hochstrasser, M. Interaction of the Doa4 deubiquitinating enzyme with the yeast 26S proteasome. Mol. Biol. Cell 10, 741–756 (1999)
Glickman, M. H. et al. A subcomplex of the proteasome regulatory particle required for ubiquitin-conjugate degradation and related to the COP9-signalosome and eIF3. Cell 94, 615–623 (1998)
Lyapina, S. et al. Promotion of NEDD-CUL1 conjugate cleavage by COP9 signalosome. Science 292, 1382–1385 (2001)
Hofmann, K. & Bucher, P. The PCI domain: a common theme in three multiprotein complexes. Trends Biochem. Sci. 23, 204–205 (1998)
Glickman, M. H., Rubin, D. M., Fried, V. A. & Finley, D. The regulatory particle of the Saccharomyces cerevisiae proteasome. Mol. Cell Biol. 18, 3149–3162 (1998)
Alberts, I. L., Nadassy, K. & Wodak, S. J. Analysis of zinc binding sites in protein crystal structures. Protein Sci. 7, 1700–1716 (1998)
Eytan, E., Armon, T., Heller, H., Beck, S. & Hershko, A. Ubiquitin C-terminal hydrolase activity associated with the 26 S protease complex. J. Biol. Chem. 268, 4668–4674 (1993)
Wilkinson, K. D. Regulation of ubiquitin-dependent processes by deubiquitinating enzymes. FASEB J. 11, 1245–1256 (1997)
Ponting, C. P., Aravind, L., Schultz, J., Bork, P. & Koonin, E. V. Eukaryotic signalling domain homologues in archaea and bacteria. Ancient ancestry and horizontal gene transfer. J. Mol. Biol. 289, 729–745 (1999)
Thick, J., Mak, Y. F., Metcalfe, J., Beatty, D. & Taylor, A. M. A gene on chromosome Xq28 associated with T-cell prolymphocytic leukemia in two patients with ataxia telangiectasia. Leukemia 8, 564–573 (1994)
Beers, E. P. & Callis, J. Utility of polyhistidine-tagged ubiquitin in the purification of ubiquitin-protein conjugates and as an affinity ligand for the purification of ubiquitin-specific hydrolases. J. Biol. Chem. 268, 21645–21649 (1993)
Hoffman, L., Pratt, G. & Rechsteiner, M. Multiple forms of the 20S multicatalytic and the 26S ubiquitin/ATP- dependent proteases from rabbit reticulocyte lysate. J. Biol. Chem. 267, 22362–22368 (1992)
Piotrowski, J. et al. Inhibition of the 26S proteasome by polyubiquitin chains synthesized to have defined lengths. J. Biol. Chem. 272, 23712–23721 (1997)
Acknowledgements
We thank D. Finley and D. Leggett for yeast strains, advice on proteasome purification, and a sample of yeast proteasomes; G. DeMartino for bovine PA700; A. Robertson for ovomucoid protein; C. Pickart for the Ub(V76)DHFR plasmid; K. Wilkinson, H. Ploegh and A. Borodovsky for UbVS, recombinant Ubp6 protein and anti-human Ubp6; C. Pickart, A. Lam and K. Wilkinson for discussions; and L. Weisman and C. Pickart for comments on the manuscript. This work was supported by a grant from the NIH and a UI Biocatalysis and Bioprocessing Fellowship to T.Y.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare that they have no competing financial interests.
Rights and permissions
About this article
Cite this article
Yao, T., Cohen, R. A cryptic protease couples deubiquitination and degradation by the proteasome. Nature 419, 403–407 (2002). https://doi.org/10.1038/nature01071
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nature01071
This article is cited by
-
Evaluation of the NRF1-proteasome axis as a therapeutic target in breast cancer
Scientific Reports (2023)
-
POH1/Rpn11/PSMD14: a journey from basic research in fission yeast to a prognostic marker and a druggable target in cancer cells
British Journal of Cancer (2022)
-
Deubiquitinating enzymes and the proteasome regulate preferential sets of ubiquitin substrates
Nature Communications (2022)
-
Two novel piperidones induce apoptosis and antiproliferative effects on human prostate and lymphoma cancer cell lines
Investigational New Drugs (2022)
-
USP15: a review of its implication in immune and inflammatory processes and tumor progression
Genes & Immunity (2021)
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.