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PMID: 16679556 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

alpha-Synuclein budding yeast model: toxicity enhanced by impaired proteasome and oxidative stress.

Journal of molecular neuroscience : MN ·Vol. 28 ·No. 2 ·2006-00-00 ·Pages 161-78

Sharma N, Brandis KA, Herrera SK, Johnson BE, Vaidya T, Shrestha R, Debburman SK

Abstract

Parkinson's disease (PD) is a common neurodegenerative disorder that results from the selective loss of midbrain dopaminergic neurons. Misfolding and aggregation of the protein alpha-synuclein, oxidative damage, and proteasomal impairment are all hypotheses for the molecular cause of this selective neurotoxicity. Here, we describe a Saccharomyces cerevisiae model to evaluate the misfolding, aggregation, and toxicity-inducing ability of wild-type alpha-synuclein and three mutants (A30P, A53T, and A30P/A53T), and we compare regulation of these properties by dysfunctional proteasomes and by oxidative stress. We found prominent localization of wild-type and A53T alpha-synuclein near the plasma membrane, supporting known in vitro lipid-binding ability. In contrast, A30P was mostly cytoplasmic, whereas A30P/A53T displayed both types of fluorescence. Surprisingly, alpha-synuclein was not toxic to several yeast strains tested. When yeast mutants for the proteasomal barrel (doa3-1) were evaluated, delayed alpha-synuclein synthesis and membrane association were observed; yeast mutant for the proteasomal cap (sen3-1) exhibited increased accumulation and aggregation of alpha-synuclein. Both sen3-1and doa3-1 mutants exhibited synthetic lethality with alpha-synuclein. When yeasts were challenged with an oxidant (hydrogen peroxide), alpha-synuclein was extremely lethal to cells that lacked manganese superoxide dismutase Mn-SOD (sod2Delta) but not to cells that lacked copper, zinc superoxide dismutase Cu,Zn-SOD (sod1Delta). Despite the toxicity, sod2Delta cells never displayed intracellular aggregates of alpha-synuclein. We suggest that the toxic alpha-synuclein species in yeast are smaller than the visible aggregates, and toxicity might involve alpha-synuclein membrane association. Thus, yeasts have emerged effective organisms for characterizing factors and mechanisms that regulate alpha-synuclein toxicity.

MeSH Terms
Animals Cell Membrane/metabolism Cysteine Endopeptidases/genetics,metabolism Humans Mutation Oxidative Stress Parkinson Disease/physiopathology Proteasome Endopeptidase Complex/metabolism Protein Folding Recombinant Fusion Proteins/genetics,metabolism Saccharomyces cerevisiae/cytology,physiology Saccharomyces cerevisiae Proteins/genetics,metabolism alpha-Synuclein/chemistry,genetics,metabolism,toxicity
Chemicals
RPN2 protein, S cerevisiae Recombinant Fusion Proteins Saccharomyces cerevisiae Proteins alpha-Synuclein Cysteine Endopeptidases PRE2 protein, S cerevisiae Proteasome Endopeptidase Complex
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Sharma Nijee
Biology Department, Lake Forest College, Lake Forest, IL 60045, USA.
Brandis Katrina A
Herrera Sara K
Johnson Brandon E
Vaidya Tulaza
Shrestha Ruja
Debburman Shubhik K
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Article Info
Journal
Journal of molecular neuroscience : MN
Abbr.
J Mol Neurosci
ISSN
0895-8696
DOI
10.1385/JMN:28:2:161
Published
2006-00-00
Pages
161-78
Language
English
Region
United States
NLM ID
9002991
Subset
IM
Grants
NINDS NIH HHS · R15 NS048508 · United States
External Links
PubMed source DOI: 10.1385/JMN:28:2:161 View journal details
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