Active site mutants in the six regulatory particle ATPases reveal multiple roles for ATP in the proteasome
Open accessAbstract
A family of ATPases resides within the regulatory particle of the proteasome. These proteins (Rpt1-Rpt6) have been proposed to mediate substrate unfolding, which may be required for translocation of substrates through the channel that leads from the regulatory particle into the proteolytic core particle. To analyze the role of ATP hydrolysis in protein breakdown at the level of the individual ATPase, we have introduced equivalent site-directed mutations into the ATPbinding motif of each RPT gene. Non-conservative substitutions of the active-site lysine were lethal in four of six cases, and conferred a strong growth defect in two cases. Thus, the ATPases are not functionally redundant, despite their multiplicity and sequence similarity. Degradation of a specific substrate can be inhibited by ATP-binding-site substitutions in many of the Rpt proteins, indicating that they co-operate in the degradation of individual substrates. The phenotypic defects of the different rpt mutants were strikingly varied. The most divergent phenotype was that of the rpt1 mutant, which was strongly growth defective despite showing no general defect in protein turnover. In addition, rpt1 was unique among the rpt mutants in displaying a G1 cell-cycle defect. Proteasomes purified from an rpt2 mutant showed a dramatic inhibition of peptidase activity, suggesting a defect in gating of the proteasome channel. In summary, ATP promotes protein breakdown by the proteasome through multiple mechanisms, as reflected by the diverse phenotypes of the rpt mutants.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
Lack of functional ATPase caused accumulation of ubiquitinated proteins and growth defects in yeast models.
“Lack of the functional ATPase caused accumulation of ubiquitinated proteins and growth defects (Gordon et al., 1993;Seeger et al., 1996;Rubin et al., 1998).”
From Depletion of 26S Proteasomes in Mouse Brain Neurons Causes Neurodegeneration and Lewy-Like Inclusions Resembling Human Pale Bodies (2008) · cites this paper as Rubin et al., 1998
In “Protein turnover in the rpt mutants” (page not provided), the paper reports growth defects and strong stabilization of two ubiquitin–proteasome substrates in three yeast mutant strains. It does not report a general accumulation of ubiquitinated proteins, and the growth-defective rpt1S mutant did not stabilize either tested substrate. (LLM, conf=0.86)
Experimental — accuracy not yet measuredPSMC1 is an essential subunit of the 26S proteasome.
“PSMC1 is an essential subunit of the 26S proteasome”
From Depletion of 26S Proteasomes in Mouse Brain Neurons Causes Neurodegeneration and Lewy-Like Inclusions Resembling Human Pale Bodies (2008) · cites this paper as Rubin et al., 1998
The Introduction (page not available) identifies the six yeast Rpt proteins as proteasomal ATPases, says they have human homologs, and reports that the yeast RPT genes are essential for vegetative growth. Our copy does not identify PSMC1 by name or establish that the human subunit itself is essential, so the claim extends beyond the evidence given. (LLM, conf=0.84)
Experimental — accuracy not yet measuredPSMC1 has a unique role in proteolysis activity of the 20S core.
“PSMC1 also has a unique role in proteolysis activity of the 20S core.”
From Depletion of 26S Proteasomes in Mouse Brain Neurons Causes Neurodegeneration and Lewy-Like Inclusions Resembling Human Pale Bodies (2008) · cites this paper as Rubin et al., 1998
In “Peptidase defect in the rpt2RF mutant” (page unavailable), the authors suggest that yeast Rpt2 has a specialized function involving either peptidase activity or peptide access to the core particle. The cited experiments concern mutant yeast proteasomes, not PSMC1 directly, and do not establish that its role is uniquely in the 20S core’s proteolytic activity. (LLM, conf=0.83)
Experimental — accuracy not yet measuredEarly S. pombe and S. cerevisiae models of proteasomal dysfunction with mutation of mts2 or rpt2 reflect the observations in the mouse model.
“The early Schizosaccharomyces pombe and Saccharomyces cerevisiae models of proteasomal dysfunction, with mutation of the homologous ATPase mts2 or rpt2, respectively, reflect the observations in our mouse model.”
From Depletion of 26S Proteasomes in Mouse Brain Neurons Causes Neurodegeneration and Lewy-Like Inclusions Resembling Human Pale Bodies (2008) · cites this paper as Rubin et al., 1998
The ‘Isolation of an rpt2R suppressor’ and ‘Protein turnover in the rpt mutants’ sections report an S. cerevisiae rpt2 mutant with impaired proteasome function. Our copy does not describe an S. pombe mts2 mutant or establish that mts2 and rpt2 are homologous. The comparison with the mouse model is the citing authors’ own interpretation, not a finding tested in this source. (LLM, conf=0.84)
Experimental — accuracy not yet measuredThe six base ATPases are functionally nonredundant.
“Because the six base ATPases are functionally nonredundant, deletion of PSMC1 would prevent hexameric ring biogenesis in the base of the 19S RP and coassembly with the 20S CP.”
From Depletion of 26S Proteasomes in Mouse Brain Neurons Causes Neurodegeneration and Lewy-Like Inclusions Resembling Human Pale Bodies (2008) · cites this paper as Rubin et al., 1998
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
No references could be resolved yet.
Research works printed but not linked: 7
- Arendt,C.S. and Hochstrasser,M. (1997) Identification of the yeast 20S proteasome catalytic centers and subunit interactions required for active-site formation. Proc. Natl Acad. Sci. USA, 94, 7156-7161.
- Glickman,M.H., Rubin,D.M., Coux,O., Wefes,I., Pfeifer,G., Cjeka,Z., Baumeister,W., Fried,V.A. and Finley,D. (1998b) A subcomplex of the proteasome regulatory particle required for ubiquitin-conjugate degradation and related to the COP9-signalosome and eIF3. Cell, in press.
- Hershko,A. and Ciechanover,A. (1998) The ubiquitin system. Annu. Rev. Biochem., in press.
- Hershko,A., Leshinsky,E., Ganoth,D. and Heller,H. (1984) ATP- dependent degradation of ubiquitin-protein conjugates. Proc. Natl Acad. Sci. USA, 81, 1619-1623.
- Lin,P. and Sherman,F. (1997) The unique hetero-oligomeric nature of the subunits in the catalytic cooperativity of the yeast Cct chaperonin complex. Proc. Natl Acad. Sci. USA, 94, 10780-10785.
- Rose,M.D., Winston,F. and Hieter,P. (1990) Methods in Yeast Genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
- Sambrook,J., Fritsch,E.F. and Maniatis,T. (1989) Molecular Cloning. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.
Cited by
Sorted by citations
Bedford, L., Hay, D., Devoy, A., Paine, S., Powe, D. G., Seth, R., Gray, T., Topham, I., Fone, K., Rezvani, N., et al. (2008) · The Journal of Neuroscience · doi:10.1523/jneurosci.2218-08.2008 · Cited by 327 · see citing works
“…of neuron-specific 26S proteasome-depleted mice To study the direct effect of impaired 26S-mediated degradation of ubiquitinated proteins in neurons, a key subunit of the 19S RP was genetically targeted, ATPase Psmc1 (Gene ID 19179) (Rubin et al., 1998; Köhler et al., 2001; Smith et al., 2007).”
BackgroundMethodInfluentialBenaroudj, N., Zwickl, P., Seemüller, E., Baumeister, W., & Goldberg, A. L. (2003) · Molecular Cell · doi:10.1016/s1097-2765(02)00775-x · Cited by 245 · see citing works
Regulated protein turnover: snapshots of the proteasome in action
Bhattacharyya, S., Yu, H., Mim, C., & Matouschek, A. (2014) · Nature Reviews Molecular Cell Biology · doi:10.1038/nrm3741 · Cited by 240 · see citing works
Küry, S., Besnard, T., Ebstein, F., Khan, T. N., Gambin, T., Douglas, J., Bacino, C. A., Craigen, W. J., Sanders, S. J., Lehmann, A., et al. (2017) · The American Journal of Human Genetics · doi:10.1016/j.ajhg.2017.01.003 · Cited by 110 · see citing works
The 19S proteasome ATPase Sug1 plays a critical role in regulating MHC class II transcription
Bhat, K. P., Turner, J. D., Myers, S. E., Cape, A. D., Ting, J. P.-Y., & Greer, S. F. (2008) · Molecular Immunology · doi:10.1016/j.molimm.2007.12.001 · Cited by 53 · see citing works
Malik, S., Shukla, A., Sen, P., & Bhaumik, S. R. (2009) · Journal of Biological Chemistry · doi:10.1074/jbc.m109.035709 · Cited by 40 · see citing works
- Authors
- Rubin, D. M., Glickman, M. H., Larsen, C. N., Dhruvakumar, S., & Finley, D.
- Affiliations
- Harvard University (US)
- Corresponding author
- Finley, Daniel
- Venue
- The EMBO Journal
- Published
- 1998-09-01
- Volume
- 17
- Issue
- 17
- Pages
- 4909-4919
- ISSN
- 0261-4189 (ISSN-L)
- 1460-2075
- PMC
- PMC1170820
- OpenAlex
- W2020462325
- Cited by
- 297
- References
- 0
- Citations by year
- 2024: 1 · 2023: 4 · 2022: 3 · 2021: 1 · 2020: 3 · 2019: 5 · 2018: 9 · 2017: 6 · 2016: 3 · 2015: 5 · 2014: 9 · 2013: 13 · 2012: 12
- FWCI
- 6.17
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- The EMBO Journal (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- American Cancer Society
- Damon Runyon Cancer Research Foundation
- National Institutes of Health (GM43601)
- National Institute of General Medical Sciences (GM43601, F32GM019359)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ubiquitin and proteasome pathways
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Heat shock proteins research
- Health Sciences › Medicine › Epidemiology › Autophagy in Disease and Therapy
- Keywords
- proteasome, ATP hydrolysis, ATP binding site, active site lysine, site-directed mutagenesis, substrate unfolding, substrate translocation, proteolytic activity, growth defects, protein degradation
- MeSH terms
- Adenosine Triphosphatases, Adenosine Triphosphate, Binding Sites, Cysteine Endopeptidases, Hydrolysis, Multienzyme Complexes, Proteins, Suppression, Genetic, Yeasts, Mutagenesis, Site-Directed, Sequence Alignment, Proteasome Endopeptidase Complex
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
