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α-synuclein fission yeast model

Concentration—Department aggregation without plasma membrane localization or toxicity

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Abstract

Despite fission yeast's history of modeling salient cellular processes, it has not yet been used to model human neurodegeneration-linked protein misfolding. Because α-synuclein misfolding and aggregation are linked to Parkinson's disease (PD), here, we report a fission yeast (Schizosaccharomyces pombe) model that evaluates α-synuclein misfolding, aggregation, and toxicity and compare these properties with those recently characterized in budding yeast (Saccharomyces cerevisiae). Wild-type α-synuclein and three mutants (A 30P, A53T, and A30P/A53T) were expressed with thiamine-repressible promoters (using vectors of increasing promoter strength: pNMT81, pNMT41, and pNMT1) to test directly in living cells the nucleation polymerization hypothesis for α-synuclein misfolding and aggregation. In support of the hypothesis, wild-type and A53T α-synuclein formed prominent intracellular cytoplasmic inclusions within fission yeast cells in a concentration- and time-dipendent manner, whereas A30P and A30P/A53T remained diffuse throughhout the cytoplasm. A53T α-synuclein for med aggregates faster than wild-type α-synuclein and at a lower α-synuclein concentration. Unexpectedly, unlike in budding yeast, wild-type and A53T α-synuclein did not target to the plasma membrane in fission yeast, not even at low α-synuclein concentrations or as a precursor step to forming aggregates. Despite α-synuclein's extensive aggregation, it was surprisingly nontoxic to fission yeast. Future genetic dissection might yield molecular insight into this protection against toxicity. We speculate that α-synuclein toxicity might be linked to its membrane binding capacity. To conclude, S. pombe and S. cerevisiae model similar yet distinct aspects of α-synuclein biology, and both organisms shed insight into α-synuclein's role in PD pathogenesis.

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  1. Biology Department, Lake Forest College, 60045, Lake Forest, IL

    Katrina A. Brandis, Isaac F. Holmes, Samantha J. England, Nijee Sharma, Lokesh Kukreja & Shubhik K. DebBurman

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  1. Katrina A. Brandis
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  2. Isaac F. Holmes
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  3. Samantha J. England
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  4. Nijee Sharma
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Correspondence to Shubhik K. DebBurman.

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Brandis, K.A., Holmes, I.F., England, S.J. et al. α-synuclein fission yeast model. J Mol Neurosci 28, 179–191 (2006). https://doi.org/10.1385/JMN:28:2:179

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