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Reciprocal Effects of α-Synuclein Overexpression and Proteasome Inhibition in Neuronal Cells and Tissue

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Abstract

Defects in the 20S/26S proteasome and conformational changes in α-synuclein (α-syn) are implicated in the development of sporadic and familial cases of PD. The objective of this study was to evaluate whether α-syn affects proteolysis by the proteasome and, reciprocally, whether proteasome inhibition affects α-syn solubility and localization. Although α-syn directly inhibited purified 20S proteasomes reversibly in vitro, its overexpression in neuroblastoma (SH-SY5Y and SK-N-BE), embryonic kidney (HEK293) cells, or mouse brain did not affect proteasome activity. Proteasome inhibition with MG132 and epoxomicin in SH-SY5Y cells failed to induce α-syn aggregation, although it increased membrane bound forms of endogenous and overexpressed wild-type, but not mutant, α-syn. Concomitantly this treatment generated cytoplasmic α-syn inclusions devoid of polyubiquitin in a small percentage of cells. The combination of proteasome inhibition with serum deprivation, which induced oxidative stress and autophagy, caused the appearance of high molecular weight α-syn species, such as those found in Lewy bodies. Our data suggest that high concentrations of α-syn do not affect proteasome function in vivo, whereas proteasome inhibition can modify synuclein solubility, most prominently under conditions of cell stress which occur during aging. These results have implications for the convergence of age-related oxidative stress and impaired protein degradation in neurodegeneration.

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Acknowledgments

The authors thank Linda Wang for technical assistance. M. D.-B. acknowledges the Ernst Schering foundation and the Charité foundation for PhD scholarships. C. D. was supported by a fellowship from Canadian Institutes of Health Research. This work was supported by operating grants to AT from the Canadian Institutes of Health Research (MOP 64373 and MOP 84501) and Parkinson Society of Canada.

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  1. Department of Medicine, Centre for Research in Neurodegenerative Diseases, University of Toronto, 6 Queen’s Park Crescent West, Toronto, ON, M5S 3H2, Canada

    Melanie Dyllick-Brenzinger, Cheryl A. D’Souza & Anurag Tandon

  2. Institut für Biochemie, Charité-Universitätsmedizin Berlin, Berlin, Germany

    Melanie Dyllick-Brenzinger, Burkhardt Dahlmann & Peter-Michael Kloetzel

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  1. Melanie Dyllick-Brenzinger
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  2. Cheryl A. D’Souza
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  5. Anurag Tandon
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Correspondence to Anurag Tandon.

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Melanie Dyllick-Brenzinger and Cheryl A. D’Souza have contributed equally to this work.

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Dyllick-Brenzinger, M., D’Souza, C.A., Dahlmann, B. et al. Reciprocal Effects of α-Synuclein Overexpression and Proteasome Inhibition in Neuronal Cells and Tissue. Neurotox Res 17, 215–227 (2010). https://doi.org/10.1007/s12640-009-9094-1

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