Alpha-Synuclein Delays Endoplasmic Reticulum (ER)-to-Golgi Transport in Mammalian Cells by Antagonizing ER/Golgi SNAREs

Authors

Nandhakumar Thayanidhi
Jared R. Helm
Deborah C. Nycz
Marbin BentleyFollow
Yingjian Liang
Jesse C. Hay, University of Montana - MissoulaFollow

Document Type

Article

Publication Title

Molecular Biology of the Cell

Publication Date

6-2010

Volume

21

Issue

11

Disciplines

Biology | Life Sciences

Abstract

Toxicity of human alpha-synuclein when expressed in simple organisms can be suppressed by overexpression of endoplasmic reticulum (ER)-to-Golgi transport machinery, suggesting that inhibition of constitutive secretion represents a fundamental cause of the toxicity. Whether similar inhibition in mammals represents a cause of familial Parkinson's disease has not been established. We tested elements of this hypothesis by expressing human alpha-synuclein in mammalian kidney and neuroendocrine cells and assessing ER-to-Golgi transport. Overexpression of wild type or the familial disease-associated A53T mutant alpha-synuclein delayed transport by up to 50%; however, A53T inhibited more potently. The secretory delay occurred at low expression levels and was not accompanied by insoluble alpha-synuclein aggregates or mistargeting of transport machinery, suggesting a direct action of soluble alpha-synuclein on trafficking proteins. Co-overexpression of ER/Golgi arginine soluble N-ethylmaleimide-sensitive factor attachment protein receptors (R-SNAREs) specifically rescued transport, indicating that alpha-synuclein antagonizes SNARE function. Ykt6 reversed alpha-synuclein inhibition much more effectively than sec22b, suggesting a possible neuroprotective role for the enigmatic high expression of ykt6 in neurons. In in vitro reconstitutions, purified alpha-synuclein A53T protein specifically inhibited COPII vesicle docking and fusion at a pre-Golgi step. Finally, soluble alpha-synuclein A53T directly bound ER/Golgi SNAREs and inhibited SNARE complex assembly, providing a potential mechanism for toxic effects in the early secretory pathway.

DOI

10.1091/mbc.E09-09-0801

Comments

© 2010, American Society For Microbiology. See original published version here.

Rights

© 2010, American Society For Microbiology. See original published version here.

Recommended Citation

Thayanidhi, Nandhakumar; Helm, Jared R.; Nycz, Deborah C.; Bentley, Marbin; Liang, Yingjian; and Hay, Jesse C., "Alpha-Synuclein Delays Endoplasmic Reticulum (ER)-to-Golgi Transport in Mammalian Cells by Antagonizing ER/Golgi SNAREs" (2010). Biological Sciences Faculty Publications. 60.
https://scholarworks.umt.edu/biosci_pubs/60