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.

  • Article
  • Published:

Tau-driven 26S proteasome impairment and cognitive dysfunction can be prevented early in disease by activating cAMP-PKA signaling

Nature Medicine volume 22, pages 46–53 (2016)Cite this article

Subjects

Abstract

The ubiquitin proteasome system (UPS) degrades misfolded proteins including those implicated in neurodegenerative diseases. We investigated the effects of tau accumulation on proteasome function in a mouse model of tauopathy and in a cross to a UPS reporter mouse (line Ub-G76V-GFP). Accumulation of insoluble tau was associated with a decrease in the peptidase activity of brain 26S proteasomes, higher levels of ubiquitinated proteins and undegraded Ub-G76V-GFP. 26S proteasomes from mice with tauopathy were physically associated with tau and were less active in hydrolyzing ubiquitinated proteins, small peptides and ATP. 26S proteasomes from normal mice incubated with recombinant oligomers or fibrils also showed lower hydrolyzing capacity in the same assays, implicating tau as a proteotoxin. Administration of an agent that activates cAMP–protein kinase A (PKA) signaling led to attenuation of proteasome dysfunction, probably through proteasome subunit phosphorylation. In vivo, this led to lower levels of aggregated tau and improvements in cognitive performance.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Tauopathy is associated with a progressive decrease in proteasome function.
Figure 2: Aggregated tau directly inhibits 26S proteasomes and associates with proteasomes in brains of mice with tauopathy.
Figure 3: Activation of PKA stimulates hydrolyzing activity of the proteasome in slices-ex vivo and in vitro.
Figure 4: Rolipram administration reduces accumulation of tau species and p62 in vivo.
Figure 5: Rolipram treatment increases proteasome function and reduces ubiquitinated protein accumulation in vivo.
Figure 6: Rolipram treatment improves cognition in rTg4510 mice with early-stage disease.

Similar content being viewed by others

References

  1. Goldberg, A.L. Protein degradation and protection against misfolded or damaged proteins. Nature 426, 895–899 (2003).

    Article  CAS  PubMed  Google Scholar 

  2. Finley, D. Recognition and processing of ubiquitin-protein conjugates by the proteasome. Annu. Rev. Biochem. 78, 477–513 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Lander, G.C. et al. Complete subunit architecture of the proteasome regulatory particle. Nature 482, 186–191 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Beck, F. et al. Near-atomic resolution structural model of the yeast 26S proteasome. Proc. Natl. Acad. Sci. USA 109, 14870–14875 (2012).

    Article  PubMed  PubMed Central  Google Scholar 

  5. Asai, M. et al. PKA rapidly enhances proteasome assembly and activity in in vivo canine hearts. J. Mol. Cell. Cardiol. 46, 452–462 (2009).

    Article  CAS  PubMed  Google Scholar 

  6. Zhang, F. et al. Proteasome function is regulated by cyclic AMP-dependent protein kinase through phosphorylation of Rpt6. J. Biol. Chem. 282, 22460–22471 (2007).

    Article  CAS  PubMed  Google Scholar 

  7. Myeku, N., Wang, H. & Figueiredo-Pereira, M.E. cAMP stimulates the ubiquitin/proteasome pathway in rat spinal cord neurons. Neurosci. Lett. 527, 126–131 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Keller, J.N., Hanni, K.B. & Markesbery, W.R. Impaired proteasome function in Alzheimer's disease. J. Neurochem. 75, 436–439 (2000).

    Article  CAS  PubMed  Google Scholar 

  9. Cripps, D. et al. Alzheimer disease-specific conformation of hyperphosphorylated paired helical filament-Tau is polyubiquitinated through Lys-48, Lys-11, and Lys-6 ubiquitin conjugation. J. Biol. Chem. 281, 10825–10838 (2006).

    Article  CAS  PubMed  Google Scholar 

  10. Thomas, S.N., Cripps, D. & Yang, A.J. Proteomic analysis of protein phosphorylation and ubiquitination in Alzheimer's disease. Methods Mol. Biol. 566, 109–121 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Morris, M. et al. Tau post-translational modifications in wild-type and human amyloid precursor protein transgenic mice. Nat. Neurosci. 18, 1183–1189 (2015).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Tai, H.C. et al. The synaptic accumulation of hyperphosphorylated tau oligomers in Alzheimer disease is associated with dysfunction of the ubiquitin-proteasome system. Am. J. Pathol. 181, 1426–1435 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Lee, M.J., Lee, J.H. & Rubinsztein, D.C. Tau degradation: the ubiquitin-proteasome system versus the autophagy-lysosome system. Prog. Neurobiol. 105, 49–59 (2013).

    Article  CAS  PubMed  Google Scholar 

  14. David, D.C. et al. Proteasomal degradation of tau protein. J. Neurochem. 83, 176–185 (2002).

    Article  CAS  PubMed  Google Scholar 

  15. Han, D.H. et al. Direct cellular delivery of human proteasomes to delay tau aggregation. Nat. Commun. 5, 5633 (2014).

    Article  CAS  PubMed  Google Scholar 

  16. Keck, S., Nitsch, R., Grune, T. & Ullrich, O. Proteasome inhibition by paired helical filament-tau in brains of patients with Alzheimer's disease. J. Neurochem. 85, 115–122 (2003).

    Article  CAS  PubMed  Google Scholar 

  17. Metcalfe, M.J., Huang, Q. & Figueiredo-Pereira, M.E. Coordination between proteasome impairment and caspase activation leading to TAU pathology: neuroprotection by cAMP. Cell Death Dis. 3, e326 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Santacruz, K. et al. Tau suppression in a neurodegenerative mouse model improves memory function. Science 309, 476–481 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Besche, H.C. & Goldberg, A.L. Affinity purification of mammalian 26S proteasomes using an ubiquitin-like domain. Methods Mol. Biol. 832, 423–432 (2012).

    Article  CAS  PubMed  Google Scholar 

  20. Peth, A., Kukushkin, N., Bossé, M. & Goldberg, A.L. Ubiquitinated proteins activate the proteasomal ATPases by binding to Usp14 or Uch37 homologs. J. Biol. Chem. 288, 7781–7790 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Verma, R., McDonald, H., Yates, J.R. III & Deshaies, R.J. Selective degradation of ubiquitinated Sic1 by purified 26S proteasome yields active S phase cyclin-Cdk. Mol. Cell 8, 439–448 (2001).

    Article  CAS  PubMed  Google Scholar 

  22. Bar-Nun, S. & Glickman, M.H. Proteasomal AAA-ATPases: structure and function. Biochim. Biophys. Acta 1823, 67–82 (2012).

    Article  CAS  PubMed  Google Scholar 

  23. Smith, D.M., Fraga, H., Reis, C., Kafri, G. & Goldberg, A.L. ATP binds to proteasomal ATPases in pairs with distinct functional effects, implying an ordered reaction cycle. Cell 144, 526–538 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Lindsten, K., Menéndez-Benito, V., Masucci, M.G. & Dantuma, N.P. A transgenic mouse model of the ubiquitin/proteasome system. Nat. Biotechnol. 21, 897–902 (2003).

    Article  CAS  PubMed  Google Scholar 

  25. Lewis, J. et al. Neurofibrillary tangles, amyotrophy and progressive motor disturbance in mice expressing mutant (P301L) tau protein. Nat. Genet. 25, 402–405 (2000).

    Article  CAS  PubMed  Google Scholar 

  26. Deriziotis, P. et al. Misfolded PrP impairs the UPS by interaction with the 20S proteasome and inhibition of substrate entry. EMBO J. 30, 3065–3077 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Wu, J.W. et al. Small misfolded Tau species are internalized via bulk endocytosis and anterogradely and retrogradely transported in neurons. J. Biol. Chem. 288, 1856–1870 (2013).

    Article  CAS  PubMed  Google Scholar 

  28. Park, S.J. et al. Resveratrol ameliorates aging-related metabolic phenotypes by inhibiting cAMP phosphodiesterases. Cell 148, 421–433 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Zhu, B. et al. Protein kinase A phosphorylation of tau-serine 214 reorganizes microtubules and disrupts the endothelial cell barrier. Am. J. Physiol. Lung Cell. Mol. Physiol. 299, L493–L501 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Schneider, A., Biernat, J., von Bergen, M., Mandelkow, E. & Mandelkow, E.M. Phosphorylation that detaches tau protein from microtubules (Ser262, Ser214) also protects it against aggregation into Alzheimer paired helical filaments. Biochemistry 38, 3549–3558 (1999).

    Article  CAS  PubMed  Google Scholar 

  31. Sadik, G. et al. Phosphorylation of tau at Ser214 mediates its interaction with 14-3-3 protein: implications for the mechanism of tau aggregation. J. Neurochem. 108, 33–43 (2009).

    Article  CAS  PubMed  Google Scholar 

  32. Lokireddy, S., Kukushkin, N.V. & Goldberg, A.L. cAMP-induced phosphorylation of 26S proteasomes on Rpn6/PSMD11 enhances their activity and the degradation of misfolded proteins. Proc. Natl. Acad. Sci. USA (in the press).

  33. Myeku, N. & Figueiredo-Pereira, M.E. Dynamics of the degradation of ubiquitinated proteins by proteasomes and autophagy: association with sequestosome 1/p62. J. Biol. Chem. 286, 22426–22440 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Cherra, S.J. III et al. Regulation of the autophagy protein LC3 by phosphorylation. J. Cell Biol. 190, 533–539 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Kandel, E.R., Dudai, Y. & Mayford, M.R. The molecular and systems biology of memory. Cell 157, 163–186 (2014).

    Article  CAS  PubMed  Google Scholar 

  36. Peth, A., Uchiki, T. & Goldberg, A.L. ATP-dependent steps in the binding of ubiquitin conjugates to the 26S proteasome that commit to degradation. Mol. Cell 40, 671–681 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Sha, Z., Peth, A. & Goldberg, A.L. Keeping proteasomes under control–a role for phosphorylation in the nucleus. Proc. Natl. Acad. Sci. USA 108, 18573–18574 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Lee, B.H. et al. Enhancement of proteasome activity by a small-molecule inhibitor of USP14. Nature 467, 179–184 (2010).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Lin, J.T. et al. Regulation of feedback between protein kinase A and the proteasome system worsens Huntington's disease. Mol. Cell. Biol. 33, 1073–1084 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Peth, A., Nathan, J.A. & Goldberg, A.L. The ATP costs and time required to degrade ubiquitinated proteins by the 26 S proteasome. J. Biol. Chem. 288, 29215–29222 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Liu, L. et al. Trans-synaptic spread of tau pathology in vivo. PLoS One 7, e31302 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. de Calignon, A. et al. Propagation of tau pathology in a model of early Alzheimer's disease. Neuron 73, 685–697 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Duff, K., Noble, W., Gaynor, K. & Matsuoka, Y. Organotypic slice cultures from transgenic mice as disease model systems. J. Mol. Neurosci. 19, 317–320 (2002).

    Article  CAS  PubMed  Google Scholar 

  44. Morris, R. Developments of a water-maze procedure for studying spatial learning in the rat. J. Neurosci. Methods 11, 47–60 (1984).

    Article  CAS  PubMed  Google Scholar 

  45. Myeku, N., Metcalfe, M.J., Huang, Q. & Figueiredo-Pereira, M. Assessment of proteasome impairment and accumulation/aggregation of ubiquitinated proteins in neuronal cultures. Methods Mol. Biol. 793, 273–296 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Saeki, Y., Isono, E. & Toh-E, A. Preparation of ubiquitinated substrates by the PY motif-insertion method for monitoring 26S proteasome activity. Methods Enzymol. 399, 215–227 (2005).

    Article  CAS  PubMed  Google Scholar 

  47. Peth, A., Besche, H.C. & Goldberg, A.L. Ubiquitinated proteins activate the proteasome by binding to Usp14/Ubp6, which causes 20S gate opening. Mol. Cell 36, 794–804 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Lam, Y.A., Huang, J.W. & Showole, O. The synthesis and proteasomal degradation of a model substrate Ub5DHFR. Methods Enzymol. 398, 379–390 (2005).

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank J. Lewis and K. Ashe for providing rTg4510 and JNPL3 mice and P. Davies for the generous gift of tau antibodies. We thank L. Liu for assisting with mice perfusion. This work was supported by grants from the US National Institute of Neurological Disorders and Stroke NS074593 (K.E.D.), CurePSP Foundation (N.M.), the US National Institute of General Medical Sciences GM051923 (to A.L.G.), the Fidelity Biosciences Research Initiative (A.L.G.) and the Multiple Myeloma Research Foundation (N.V.K.).

Author information

Authors and Affiliations

  1. Department of Pathology and Cell Biology, Taub Institute for Alzheimer's Disease Research, Columbia University, New York, New York, USA

    Natura Myeku, Catherine L Clelland, Sheina Emrani, Wai Haung Yu & Karen E Duff

  2. Department of Cell Biology, Harvard Medical School, Boston, Massachusetts, USA

    Nikolay V Kukushkin & Alfred L Goldberg

  3. Division of Integrative Neuroscience in the Department of Psychiatry, New York State Psychiatric Institute, New York, New York, USA

    Karen E Duff

Authors
  1. Natura Myeku
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Catherine L Clelland
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sheina Emrani
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Nikolay V Kukushkin
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Wai Haung Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Alfred L Goldberg
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Karen E Duff
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

N.M. performed all experiments except for the degradation assay of ubiquitinated proteins. C.L.C. performed STATA analyses for behavioral studies; S.E. assisted with IHC and water maze experiments; N.V.K. performed degradation assays. N.M., W.H.Y., A.L.G. and K.E.D. designed the studies. N.M., A.L.G. and K.E.D. wrote the manuscript. C.L.C., N.V.K. and W.H.Y. contributed to manuscript preparation. W.H.Y. contributed transfected cell lines. All authors reviewed and commented on the manuscript.

Corresponding author

Correspondence to Karen E Duff.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–12 and Supplementary Data (PDF 3617 kb)

Source data

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Myeku, N., Clelland, C., Emrani, S. et al. Tau-driven 26S proteasome impairment and cognitive dysfunction can be prevented early in disease by activating cAMP-PKA signaling. Nat Med 22, 46–53 (2016). https://doi.org/10.1038/nm.4011

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nm.4011

This article is cited by

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