Abstract
Impairment of the ubiquitin/proteasome system has been proposed to play a role in neurodegenerative disorders such as Alzheimer and Parkinson diseases. Although recent studies confirmed that some disease-related proteins block proteasomal degradation, and despite the existence of excellent animal models of both diseases, in vivo data about the system are lacking. We have developed a model for in vivo analysis of the ubiquitin/proteasome system by generating mouse strains transgenic for a green fluorescent protein (GFP) reporter carrying a constitutively active degradation signal. Administration of proteasome inhibitors to the transgenic animals resulted in a substantial accumulation of GFP in multiple tissues, confirming the in vivo functionality of the reporter. Moreover, accumulation of the reporter was induced in primary neurons by UBB+1, an aberrant ubiquitin found in Alzheimer disease. These transgenic animals provide a tool for monitoring the status of the ubiquitin/proteasome system in physiologic or pathologic conditions.
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References
Sherman, M.Y. & Goldberg, A.L. Cellular defenses against unfolded proteins: a cell biologist thinks about neurodegenerative diseases. Neuron 29, 15–32 (2001).
DeSalle, L.M. & Pagano, M. Regulation of the G1 to S transition by the ubiquitin pathway. FEBS Lett. 490, 179–189 (2001).
Karin, M. & Ben-Neriah, Y. Phosphorylation meets ubiquitination: the control of NF-κB activity. Annu. Rev. Immunol. 18, 621–663 (2000).
Baumeister, W., Walz, J., Zuhl, F. & Seemuller, E. The proteasome: paradigm of a self-compartmentalizing protease. Cell 92, 367–380 (1998).
Hershko, A. & Ciechanover, A. The ubiquitin system. Annu. Rev. Biochem. 67, 425–479 (1998).
Bence, N.F., Sampat, R.M. & Kopito, R.R. Impairment of the ubiquitin-proteasome system by protein aggregation. Science 292, 1552–1555 (2001).
Cummings, C.J. et al. Chaperone suppression of aggregation and altered subcellular proteasome localization imply protein misfolding in SCA1. Nat. Genet. 19, 148–154 (1998).
Lam, Y.A. et al. Inhibition of the ubiquitin-proteasome system in Alzheimer's disease. Proc. Natl. Acad. Sci. USA 97, 9902–9906 (2000).
Lindsten, K. et al. Mutant ubiquitin found in neurodegenerative disorders is a ubiquitin fusion degradation substrate that blocks proteasomal degradation. J. Cell Biol. 157, 417–427 (2002).
Mizuno, Y., Hattori, N., Mori, H., Suzuki, T. & Tanaka, K. Parkin and Parkinson's disease. Curr. Opin. Neurol. 14, 477–482 (2001).
Carrell, R.W. & Lomas, D.A. Conformational disease. Lancet 350, 134–138 (1997).
Muchowski, P.J. Protein misfolding, amyloid formation, and neurodegeneration: a critical role for molecular chaperones? Neuron 35, 9–12 (2002).
Soto, C. Protein misfolding and disease; protein refolding and therapy. FEBS Lett. 498, 204–207 (2001).
Verhoef, L.G., Lindsten, K., Masucci, M.G. & Dantuma, N.P. Aggregate formation inhibits proteasomal degradation of polyglutamine proteins. Hum. Mol. Genet. 11, 2689–2700 (2002).
Floyd, J.A. & Hamilton, B.A. Intranuclear inclusions and the ubiquitin-proteasome pathway: digestion of a red herring? Neuron 24, 765–766 (1999).
Saudou, F., Finkbeiner, S., Devys, D. & Greenberg, M.E. Huntingtin acts in the nucleus to induce apoptosis but death does not correlate with the formation of intranuclear inclusions. Cell 95, 55–66 (1998).
Watase, K. et al. A long CAG repeat in the mouse Sca1 locus replicates SCA1 features and reveals the impact of protein solubility on selective neurodegeneration. Neuron 34, 905–919 (2002).
Ryan, K.M., Phillips, A.C. & Vousden, K.H. Regulation and function of the p53 tumor suppressor protein. Curr. Opin. Cell. Biol. 13, 332–337 (2001).
Slingerland, J. & Pagano, M. Regulation of the cdk inhibitor p27 and its deregulation in cancer. J. Cell Physiol. 183, 10–17 (2000).
Meng, L. et al. Epoxomicin, a potent and selective proteasome inhibitor, exhibits in vivo antiinflammatory activity. Proc. Natl. Acad. Sci. USA 96, 10403–10408 (1999).
Meng, L., Kwok, B.H., Sin, N. & Crews, C.M. Eponemycin exerts its antitumor effect through the inhibition of proteasome function. Cancer Res. 59, 2798–2801 (1999).
Aghajanian, C. et al. A phase I trial of the novel proteasome inhibitor PS341 in advanced solid tumor malignancies. Clin. Cancer Res. 8, 2505–2511 (2002).
Pati, S. et al. Antitumorigenic effects of HIV protease inhibitor ritonavir: inhibition of Kaposi sarcoma. Blood 99, 3771–3779 (2002).
Sgadari, C. et al. HIV protease inhibitors are potent anti-angiogenic molecules and promote regression of Kaposi sarcoma. Nat. Med. 8, 225–232 (2002).
Hosseini, H. et al. Protection against experimental autoimmune encephalomyelitis by a proteasome modulator. J. Neurobiol. 118, 233–244 (2001).
Zollner, T.M. et al. Proteasome inhibition reduces superantigen-mediated T cell activation and the severity of psoriasis in a SCID-hu model. J. Clin. Invest. 109, 671–679 (2002).
Luo, H. et al. A proteasome inhibitor effectively prevents mouse heart allograft rejection. Transplantation 72, 196–202 (2001).
Rock, K.L. & Goldberg, A.L. Degradation of cell proteins and the generation of MHC class I–presented peptides. Annu. Rev. Immunol. 17, 739–779 (1999).
Rubinsztein, D.C. Lessons from animal models of Huntington's disease. Trends Genet. 18, 202–209 (2002).
Wong, P.C., Cai, H., Borchelt, D.R. & Price, D.L. Genetically engineered mouse models of neurodegenerative diseases. Nat. Neurosci. 5, 633–639 (2002).
Dantuma, N.P., Lindsten, K., Glas, R., Jellne, M. & Masucci, M.G. Short-lived green fluorescent proteins for quantification of ubiquitin/proteasome-dependent proteolysis in living cells. Nat. Biotechnol. 18, 538–543 (2000).
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).
Okabe, M., Ikawa, M., Kominami, K., Nakanishi, T. & Nishimune, Y. 'Green mice' as a source of ubiquitous green cells. FEBS Lett. 407, 313–319 (1997).
Jensen, T.J. et al. Multiple proteolytic systems, including the proteasome, contribute to CFTR processing. Cell 83, 129–135 (1995).
McCormack, T. et al. Active site-directed inhibitors of Rhodococcus 20 S proteasome. Kinetics and mechanism. J. Biol. Chem. 272, 26103–26109 (1997).
Bogyo, M. et al. Covalent modification of the active site threonine of proteasomal beta subunits and the Escherichia coli homolog HslV by a new class of inhibitors. Proc. Natl. Acad. Sci. USA 94, 6629–6634 (1997).
Kisselev, A.F. & Goldberg, A.L. Proteasome inhibitors: from research tools to drug candidates. Chem. Biol. 8, 739–758 (2001).
Myung, J., Kim, K.B., Lindsten, K., Dantuma, N.P. & Crews, C.M. Lack of proteasome active site allostery as revealed by subunit-specific inhibitors. Mol. Cell 7, 411–420 (2001).
van Leeuwen, F.W. et al. Frameshift mutants of β-amyloid precursor protein and ubiquitin-B in Alzheimer's and Down patients. Science 279, 242–247 (1998).
Perutz, M.F. & Windle, A.H. Cause of neural death in neurodegenerative diseases attributable to expansion of glutamine repeats. Nature 412, 143–144 (2001).
Sisodia, S.S. Nuclear inclusions in glutamine repeat disorders: are they pernicious, coincidental, or beneficial? Cell 95, 1–4 (1998).
Lindsten, K. & Dantuma, N.P. Monitoring the ubiquitin/proteasome system in conformational diseases. Ageing Res. Rev. in the press.
French, B.A. et al. Aggresome formation in liver cells in response to different toxic mechanisms: role of the ubiquitin-proteasome pathway and the frameshift mutant of ubiquitin. Exp. Mol. Pathol. 71, 241–246 (2001).
Denk, H., Stumptner, C. & Zatloukal, K. Mallory bodies revisited. J. Hepatol. 32, 689–702 (2000).
Dallaporta, B. et al. Proteasome activation as a critical event of thymocyte apoptosis. Cell Death Differ. 7, 368–373 (2000).
Yang, Y., Fang, S., Jensen, J.P., Weissman, A.M. & Ashwell, J.D. Ubiquitin protein ligase activity of IAPs and their degradation in proteasomes in response to apoptotic stimuli. Science 288, 874–877 (2000).
Naldini, L. et al. In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector. Science 272, 263–267 (1996).
Acknowledgements
We thank Marianne Jellne, Ros-Mari Johansson, Margareta Hagelin and Maj-Britt Alter for technical assistance, and Hidde Ploegh, Eva Backström, Luigi Naldini, Fred van Leeuwen, Elly Hol, David Fischer, Björn Rozell, Johan Brask, Xu Hong, Jelena Petrovic, Petter Höglund and Jacques Neefjes for reagents, advice and technical help. Pronuclear injections were performed by the Karolinska Center for Transgene Technologies, and pathology examinations were performed by the Mouse Pathology Core Facility of the Karolinska Institute. This work was supported by grants awarded by the Swedish Research Council (N.P.D.), Swedish Cancer Society (N.P.D., M.G.M.), the Swedish Foundation of Strategy Research (M.G.M.), the Swedish Alzheimer Foundation (N.P.D.) and the Karolinska Institute. N.P.D. is supported by a fellowship awarded by the Swedish Research Council.
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Lindsten, K., Menéndez-Benito, V., Masucci, M. et al. A transgenic mouse model of the ubiquitin/proteasome system. Nat Biotechnol 21, 897–902 (2003). https://doi.org/10.1038/nbt851
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DOI: https://doi.org/10.1038/nbt851
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