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:

Synphilin-1 associates with α-synuclein and promotes the formation of cytosolic inclusions

Nature Genetics volume 22, pages 110–114 (1999)Cite this article

Abstract

Parkinson disease (PD) is a neurodegenerative disease characterized by tremor, bradykinesia, rigidity and postural instability. Post-mortem examination shows loss of neurons and Lewy bodies, which are cytoplasmic eosinophilic inclusions, in the substantia nigra and other brain regions1,2. A few families have PD caused by mutations (A53T or A30P) in the gene SNCA (encoding α-synuclein; refs 3, 4, 5). α-synuclein is present in Lewy bodies of patients with sporadic PD (Refs 6,7), suggesting that α-synuclein may be involved in the pathogenesis of PD. It is unknown how α-synuclein contributes to the cellular and biochemical mechanisms of PD, and its normal functions and biochemical properties are poorly understood8,9,10. To determine the protein-interaction partners of α-synuclein, we performed a yeast two-hybrid screen. We identified a novel interacting protein, which we term synphilin-1 (encoded by the gene SNCAIP). We found that α-synuclein interacts in vivo with synphilin-1 in neurons. Co-transfection of both proteins (but not control proteins) in HEK 293 cells yields cytoplasmic eosinophilic inclusions.

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

Access options

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

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

Figure 1: Association of α-synuclein with synphilin-1 in yeast.
Figure 2: Synphilin-1 ORF.
Figure 3: Interaction of α-synuclein and synphilin-1.
Figure 4: Expression pattern of synphilin-1 and co-localization with α-synuclein.
Figure 5: Formation of cytoplasmic inclusions by co-transfection of constructs encoding NAC and synphilin-1.

Similar content being viewed by others

Accession codes

Accessions

GenBank/EMBL/DDBJ

References

  1. Forno, L.S. Neuropathology of Parkinson's disease. J. Neuropathol. Exp. Neurol. 55, 259–272 (1996).

    Article  CAS  Google Scholar 

  2. Pollanen, M.S., Dickson, D.W. & Bergeron, C. Pathology and biology of the Lewy body. J. Neuropathol. Exp. Neurol. 52, 183–191 (1993).

    Article  CAS  Google Scholar 

  3. Nussbaum, R.L. & Polymeropoulos, M.H. Genetics of Parkinson's disease. Hum. Mol. Genet. 6, 1687–1691 (1997).

    Article  CAS  Google Scholar 

  4. Polymeropoulos, M.H. et al. Mutation in the α-synuclein gene identified in families with Parkinson's disease. Science 276, 2045–2047 (1997).

    Article  CAS  Google Scholar 

  5. Kruger, R. et al. Ala30Pro mutation in the gene encoding α-synuclein in Parkinson's disease. Nature Genet. 18, 106–108 (1998).

    Article  CAS  Google Scholar 

  6. Spillantini, M.G. et al. α-synuclein in Lewy bodies. Nature 388, 839–840 (1997).

    Article  CAS  Google Scholar 

  7. Spillantini, M.G. et al. α-synuclein in filamentous inclusions of Lewy bodies from Parkinson's disease and dementia with Lewy bodies. Proc. Natl Acad. Sci. USA 95, 6469–6473 (1998).

    Article  CAS  Google Scholar 

  8. Maroteaux, L., Campanelli, J.T. & Scheller, R.H. Synuclein: a neuron-specific protein localized to the nucleus and presynaptic nerve terminal. J. Neurosci. 8, 2804–2815 (1988).

    Article  CAS  Google Scholar 

  9. Jenco, J.M., Rawlingson, A., Daniels, B. & Morris, A.J. Regulation of phospholipase D2: selective inhibition of mammalian phospholipase D isoenzymes by α- and β-synucleins. Biochemistry 37, 4901–4909 (1998).

    Article  CAS  Google Scholar 

  10. Clayton, D.F. & George, J.M. The synucleins: a family of proteins involved in synaptic function, plasticity, neurodegeneration and disease. Trends Neurosci. 21, 249–254 (1998).

    Article  CAS  Google Scholar 

  11. Chevray, P.M. & Nathans, D. Protein interaction cloning in yeast: identification of mammalian proteins that react with leucine zipper of Jun. Proc. Natl Acad. Sci. USA 89, 5789–5793 (1992).

    Article  CAS  Google Scholar 

  12. Lux, S.E., John, K.M. & Bennett, V. Analysis of cDNA for human erythrocyte ankyrin indicates a repeated structure with homology to tissue-differentiation and cell-cycle control protein. Nature 344, 36–42 (1990).

    Article  CAS  Google Scholar 

  13. Lupas, A. Prediction and analysis of coiled-coil structures. Methods Enzymol. 266, 513–525 (1996).

    Article  CAS  Google Scholar 

  14. Engelender, S. et al. Huntingtin associated protein 1 (HAP1) interacts with dynactin p150Glued and other cytoskeletal related proteins. Hum. Mol. Genet. 13, 2205–2212 (1997).

    Article  Google Scholar 

  15. Withers, G.S. et al. Delayed localization of synelfin (synuclein, NACP) to presynaptic terminals in cultured rat hippocampal neurons. Dev. Brain Res. 99, 87–94 (1997).

    Article  CAS  Google Scholar 

  16. Kuzuhara, S. et al. Lewy bodies are ubiquitinated. A light and electron microscopic immunocytochemical study. Acta Neuropathol. 75, 345–353 (1988).

    Article  CAS  Google Scholar 

  17. Baba, M. et al. Aggregation of α-synuclein in Lewy bodies of sporadic Parkinson's disease and dementia with Lewy bodies. Am. J. Pathol. 152, 879–884 (1998).

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Selkoe, D.J. The cell biology of β-amyloid precursor protein and presenilin in Alzheimer's disease. Trends Cell Biol. 8, 447–453 (1998).

    Article  CAS  Google Scholar 

  19. Kim, T.W., Pettingell, W.H., Jung, Y.K., Kovacs, D.M. & Tanzi, R.E. Alternative cleavage of Alzheimer-associated presenilins during apoptosis by caspase-3 family protease. Science 277, 373–376 (1997).

    Article  CAS  Google Scholar 

  20. Ross, C.A. Intranuclear neuronal inclusions: a common pathogenic mechanism for glutamine-repeat neurodegenerative diseases? Neuron 19, 1147–1150 (1997).

    Article  CAS  Google Scholar 

  21. Conway, K.A., Harper, J.D. & Lansburry, P.T. Accelerated in vitro fibril formation by a mutant α-synuclein linked to early-onset Parkinson disease. Nature Med. 4, 1318–1320 (1998).

    Article  CAS  Google Scholar 

  22. El-Agnaf, O.M.A., Jakes, R., Curran, M.D. & Wallace, A. Effects of the mutations Ala30 to Pro and Ala53 to Thr on the physical and morphological properties of α-synuclein protein implicated in Parkinson's disease. FEBS Lett. 440, 67–70 (1998).

    Article  CAS  Google Scholar 

  23. Li, X.-J. et al. A huntingtin-associated protein enriched in brain with implications for pathology. Nature 378, 398–402 (1995).

    Article  CAS  Google Scholar 

  24. Dawson, V.L. et al. Mechanisms of nitric oxide-mediated neurotoxicity in primary brain cultures. J. Neurosci. 13, 2651–2661.

  25. Uéda, K. et al. Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease. Proc. Natl Acad. Sci. USA 90, 11282–11286 (1993).

    Article  Google Scholar 

  26. Iwai, A., Yoshimoto, M., Masliah, E. & Saitoh, T. Non-Aβ component of Alzheimer's disease amyloid (NAC) is amyloidogenic. Biochemistry 34, 10139–10145 (1995).

    Article  CAS  Google Scholar 

  27. Han, H., Weinreb, P.H. & Lansbury, P.T. Jr The core Alzheimer's peptide NAC forms amyloid fibrils which seed and are seeded by β-amyloid: is NAC a common trigger or target in neurodegenerative disease? Chem. Biol. 2, 163–169 (1995).

    Article  CAS  Google Scholar 

  28. Heintz, N. & Zoghbi, H. α-synuclein—a link between Parkinson and Alzheimer diseases? Nature Genet. 16, 325–327 (1997).

    Article  CAS  Google Scholar 

  29. Nakajo, S. et al. Purification and characterization of a novel brain-specific 14-kDa protein. J. Neurochem. 55, 2031–2038 (1990).

    Article  CAS  Google Scholar 

  30. Jakes, R., Spillantini, M.G. & Goedert, M. Identification of two distinct synucleins from human brain. FEBS Lett. 345, 27–32 (1994).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank G.L. Rudow and C. Zhang for technical support, B.E. Hoffman for providing the primary cultures and M. Delanoy for the assistance with confocal microscopy. This work was supported by a grant from the Parkinson's Disease Foundation, a Pew Fellows Award to S.E., NINDS NS38377 and NS16375, the DeVelbiss fund and a bequest from Sar and Brita Levitan. X.G. is supported by the Herbert Friedburg Fellowship and T.M.D. and V.L.D. are supported by the Edward D. and Anna Mitchell Family Foundation.

Author information

Authors and Affiliations

  1. Division of Neurobiology, Department of Psychiatry, Johns Hopkins University School of Medicine,

    Simone Engelender, Zachary Kaminsky, Alan H. Sharp, Ravi K. Amaravi, John J. Kleiderlein, Russell L. Margolis & Christopher A. Ross

  2. Department of Neurology, Johns Hopkins University, Baltimore, 21205, Maryland, USA

    Xin Guo, Anthony A. Lanahan, Paul F. Worley, Valina L. Dawson & Ted M. Dawson

  3. Division of Neuropathology, Johns Hopkins University, Baltimore, 21205, Maryland, USA

    Juan C. Troncoso

  4. Department of Neuroscience, Johns Hopkins University, Baltimore, 21205, Maryland, USA

    Anthony A. Lanahan, Paul F. Worley, Valina L. Dawson, Ted M. Dawson & Christopher A. Ross

  5. Program in Cellular and Molecular Medicine, Johns Hopkins University, Baltimore, 21205, Maryland, USA

    Christopher A. Ross

Authors
  1. Simone Engelender
    View author publications

    You can also search for this author inPubMed Google Scholar

  2. Zachary Kaminsky
    View author publications

    You can also search for this author inPubMed Google Scholar

  3. Xin Guo
    View author publications

    You can also search for this author inPubMed Google Scholar

  4. Alan H. Sharp
    View author publications

    You can also search for this author inPubMed Google Scholar

  5. Ravi K. Amaravi
    View author publications

    You can also search for this author inPubMed Google Scholar

  6. John J. Kleiderlein
    View author publications

    You can also search for this author inPubMed Google Scholar

  7. Russell L. Margolis
    View author publications

    You can also search for this author inPubMed Google Scholar

  8. Juan C. Troncoso
    View author publications

    You can also search for this author inPubMed Google Scholar

  9. Anthony A. Lanahan
    View author publications

    You can also search for this author inPubMed Google Scholar

  10. Paul F. Worley
    View author publications

    You can also search for this author inPubMed Google Scholar

  11. Valina L. Dawson
    View author publications

    You can also search for this author inPubMed Google Scholar

  12. Ted M. Dawson
    View author publications

    You can also search for this author inPubMed Google Scholar

  13. Christopher A. Ross
    View author publications

    You can also search for this author inPubMed Google Scholar

Corresponding author

Correspondence to Christopher A. Ross.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Engelender, S., Kaminsky, Z., Guo, X. et al. Synphilin-1 associates with α-synuclein and promotes the formation of cytosolic inclusions. Nat Genet 22, 110–114 (1999). https://doi.org/10.1038/8820

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/8820

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