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.

  • Letter
  • Published:

Cytosolic chaperonin prevents polyglutamine toxicity with altering the aggregation state

Nature Cell Biology volume 8, pages 1163–1169 (2006)Cite this article

Abstract

Polyglutamine (polyQ)-expansion proteins cause neurodegenerative disorders including Huntington's disease, Kennedy's disease and various ataxias. The cytotoxicity of these proteins is associated with the formation of aggregates or other conformationally toxic species. Here, we show that the cytosolic chaperonin CCT (also known as TRiC) can alter the course of aggregation and cytotoxicity of huntingtin (Htt)–polyQ proteins in mammalian cells. Disruption of the CCT complex by RNAi-mediated knockdown enhanced Htt–polyQ aggregate formation and cellular toxicity. Analysis of the aggregation states of the Htt–polyQ proteins by fluorescence correlation spectroscopy revealed that CCT depletion results in the appearance of soluble Htt–polyQ aggregates. Similarly, overexpression of all eight subunits of CCT suppressed Htt aggregation and neuronal cell death. These results indicate that CCT has an essential role in protecting against the cytotoxicity of polyQ proteins by affecting the course of aggregation.

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: Knockdown of CCT subunits stimulates aggregate formation of expanded polyQ repeats transiently expressed in HEK293 cells.
Figure 2: CCT is required for preventing Htt aggregation in stable transformants of HeLa cells.
Figure 3: Depletion of CCT subunits stimulates formation of soluble aggregates of polyQ-expansion proteins.
Figure 4: CCT prevents Htt toxicity and aggregation in neuronal cells.

Similar content being viewed by others

References

  1. Ross, C. A. Polyglutamine pathogenesis: emergence of unifying mechanisms for Huntington's disease and related disorders. Neuron 35, 819–822 (2002).

    Article  CAS  Google Scholar 

  2. Gatchel, J. R. & Zoghbi, H. Y. Diseases of unstable repeat expansion: mechanisms and common principles. Nature Rev. Genet. 6, 743–755 (2005).

    Article  CAS  Google Scholar 

  3. The Huntington's Disease Collaborative Research Group. A novel gene containing a trinucleotide repeat that is expanded and unstable on Huntington's disease chromosomes. Cell 72, 971–983 (1993).

  4. DiFiglia, M. et al. Huntingtin is a cytoplasmic protein associated with vesicles in human and rat brain neurons. Neuron 14, 1075–1081 (1995).

    Article  CAS  Google Scholar 

  5. Davies, S. W. et al. Formation of neuronal intranuclear inclusions underlies the neurological dysfunction in mice transgenic for the HD mutation. Cell 90, 537–548 (1997).

    Article  CAS  Google Scholar 

  6. Scherzinger, E. et al. Huntingtin-encoded polyglutamine expansions form amyloid-like protein aggregates in vitro and in vivo. Cell 90, 549–558 (1997).

    Article  CAS  Google Scholar 

  7. Gidalevitz, T., Ben-Zvi, A., Ho, K. H., Brignull, H. R. & Morimoto, R. I. Progressive disruption of cellular protein folding in models of polyglutamine diseases. Science 311, 1471–1474 (2006).

    Article  CAS  Google Scholar 

  8. Perutz, M. F., Finch, J. T., Berriman, J. & Lesk, A. Amyloid fibers are water-filled nanotubes. Proc. Natl Acad. Sci. USA 99, 5591–5595 (2002).

    Article  CAS  Google Scholar 

  9. Young, J. C., Agashe, V. R., Siegers, K. & Hartl, F. U. Pathways of chaperone-mediated protein folding in the cytosol. Nature Rev. Mol. Cell Biol. 5, 781–791 (2004).

    Article  CAS  Google Scholar 

  10. Muchowski, P. J. & Wacker, J. L. Modulation of neurodegeneration by molecular chaperones. Nature Rev. Neurosci. 6, 11–22 (2005).

    Article  CAS  Google Scholar 

  11. Fenton, W. A. & Horwich, A. L. Chaperonin-mediated protein folding: fate of substrate polypeptide. Q. Rev. Biophys. 36, 229–256 (2003).

    Article  CAS  Google Scholar 

  12. Spiess, C., Meyer, A. S., Reissmann, S. & Frydman, J. Mechanism of the eukaryotic chaperonin: protein folding in the chamber of secrets. Trends Cell Biol. 14, 598–604 (2004).

    Article  CAS  Google Scholar 

  13. Frydman, J. & Hartl, F. U. Principles of chaperone-assisted protein folding: differences between in vitro and in vivo mechanisms. Science 272, 1497–1502 (1996).

    Article  CAS  Google Scholar 

  14. Farr, G. W., Scharl, E. C., Schumacher, R. J., Sondek, S. & Horwich, A. L. Chaperonin-mediated folding in the eukaryotic cytosol proceeds through rounds of release of native and nonnative forms. Cell 89, 927–937 (1997).

    Article  CAS  Google Scholar 

  15. Kubota, H., Hynes, G., Carne, A., Ashworth, A. & Willison, K. Identification of six Tcp-1-related genes encoding divergent subunits of the TCP-1-containing chaperonin. Curr. Biol. 4, 89–99 (1994).

    Article  CAS  Google Scholar 

  16. Llorca, O. et al. Eukaryotic chaperonin CCT stabilizes actin and tubulin folding intermediates in open quasi-native conformations. EMBO J. 19, 5971–5979 (2000).

    Article  CAS  Google Scholar 

  17. Meyer, A. S. et al. Closing the folding chamber of the eukaryotic chaperonin requires the transition state of ATP hydrolysis. Cell 113, 369–381 (2003).

    Article  CAS  Google Scholar 

  18. Nollen, E. A. et al. Genome-wide RNA interference screen identifies previously undescribed regulators of polyglutamine aggregation. Proc. Natl Acad. Sci. USA 101, 6403–6408 (2004).

    Article  CAS  Google Scholar 

  19. Kubota, S., Kubota, H. & Nagata, K. Cytosolic chaperonin protects folding intermediates of Gβ by recognizing hydrophobic β-strands. Proc. Natl Acad. Sci. USA 103, 8360–8365 (2006).

    Article  CAS  Google Scholar 

  20. Holmberg, C. I., Staniszewski, K. E., Mensah, K. N., Matouschek, A. & Morimoto, R. I. Inefficient degradation of truncated polyglutamine proteins by the proteasome. EMBO J. 23, 4307–4318 (2004).

    Article  CAS  Google Scholar 

  21. Matsumoto, G., Kim, S. & Morimoto, R. I. Huntingtin and mutant SOD1 form aggregate structures with distinct molecular properties in human cells. J. Biol. Chem. 281, 4477–4485 (2005).

    Article  Google Scholar 

  22. Muchowski, P. J. et al. Hsp70 and hsp40 chaperones can inhibit self-assembly of polyglutamine proteins into amyloid-like fibrils. Proc. Natl Acad. Sci. USA 97, 7841–7846 (2000).

    Article  CAS  Google Scholar 

  23. Schaffar, G. et al. Cellular toxicity of polyglutamine expansion proteins: mechanism of transcription factor deactivation. Mol. Cell 15, 95–105 (2004).

    Article  CAS  Google Scholar 

  24. Kim, S., Nollen, E. A., Kitagawa, K., Bindokas, V. P. & Morimoto, R. I. Polyglutamine protein aggregates are dynamic. Nature Cell. Biol. 4, 826–831 (2002).

    Article  CAS  Google Scholar 

  25. Rigler, R., Mets, U., Widengren, J. & Kask, P. Fluorescence correlation spectroscopy with high count rate and low background: analysis of translational diffusion. Eur. Biophys. J. 22, 169–175 (1993).

    Article  CAS  Google Scholar 

  26. Terada, S., Kinjo, M. & Hirokawa, N. Oligomeric tubulin in large transporting complex is transported via kinesin in squid giant axons. Cell 103, 141–155 (2000).

    Article  CAS  Google Scholar 

  27. Feldman, D. E., Spiess, C., Howard, D. E. & Frydman, J. Tumorigenic mutations in VHL disrupt folding in vivo by interfering with chaperonin binding. Mol. Cell 12, 1213–1224 (2003).

    Article  CAS  Google Scholar 

  28. Siegers, K. et al. TRiC/CCT cooperates with different upstream chaperones in the folding of distinct protein classes. EMBO J. 22, 5230–5240 (2003).

    Article  CAS  Google Scholar 

  29. Campbell, R. E. et al. A monomeric red fluorescent protein. Proc. Natl Acad. Sci. USA 99, 7877–7882 (2002).

    Article  CAS  Google Scholar 

  30. Yokota, S., Yanagi, H., Yura, T. & Kubota, H. Cytosolic chaperonin is up-regulated during cell growth. Preferential expression and binding to tubulin at G(1)/S transition through early S phase. J. Biol. Chem. 274, 37070–37078 (1999).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank M. Kadota for technical assistance and R. Tsien for providing the mRFP1 plasmid. We are grateful to I. Wada, T. Haraguchi, H. Taguchi, Y. Nagai, T. Shimi, K. Kabayama, K. Saito and H. Fujii for helpful discussion. G.M. was supported by a Human Frontiers Fellowship at Northwestern University and R.I.M. is supported by grants from the National Institutes of Health (National Institute for Medical Science, NIGMS; National Institute for Neurological Diseases and Stroke, NINDS; and National Institute for Aging, NIA), the Huntington Disease Coalition for the Cure, and the Daniel F. and Ada L. Rice Foundation. A.K. and C.P. were supported by fellowships from the Japan Society for Promotion of Science. H.K., M.K. and K.N. were supported by the Grants-in-Aid for Scientific Research programmes in Japan.

Author information

Authors and Affiliations

  1. Department of Molecular and Cellular Biology and CREST/JST, Institute for Frontier Medical Sciences, Kyoto University, 53 Shogoin Kawahara-cho, Sakyo-ku, 606-8397, Kyoto, Japan

    Akira Kitamura, Hiroshi Kubota, Shoshiro Hirayama & Kazuhiro Nagata

  2. Laboratory of Supramolecular Physics, Research Institute for Electronic Science, Hokkaido University, N12W6, Kita-ku, 060-0812, Sapporo, Japan

    Chan-Gi Pack, Yasuo Takahashi & Masataka Kinjo

  3. Department of Biochemistry, Molecular Biology and Cell Biology, Rice Institute for Biomedical Research, Northwestern University, Evanston, 60208, IL, USA

    Gen Matsumoto & Richard I. Morimoto

  4. Olympus Corporation, 2-3 Kuboyama-cho, Hachioji-shi, Tokyo, 192-8512, Japan

    Yasuo Takahashi

  5. Nuclear Function and Dynamics Unit, HMRO, School of Medicine, Kyoto University, Yoshida Konoe-cho, Sakyo-ku, 606-8501, Kyoto, Japan

    Hiroshi Kimura

Authors
  1. Akira Kitamura
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hiroshi Kubota
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Chan-Gi Pack
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Gen Matsumoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Shoshiro Hirayama
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yasuo Takahashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Hiroshi Kimura
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Masataka Kinjo
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Richard I. Morimoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Kazuhiro Nagata
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hiroshi Kubota.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

Supplementary figures S1, S2, S3, S4 and S5. (PDF 896 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kitamura, A., Kubota, H., Pack, CG. et al. Cytosolic chaperonin prevents polyglutamine toxicity with altering the aggregation state. Nat Cell Biol 8, 1163–1169 (2006). https://doi.org/10.1038/ncb1478

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ncb1478

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