α-synuclein fission yeast model -: Concentration-dependent aggregation without plasma membrane localization or toxicity
α-synuclein fission yeast model -: Concentration-dependent aggregation without plasma membrane localization or toxicity
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DOI：
10.1385/jmn:28:2:179
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发表时间：
2006-01-01
影响因子：
3.1
通讯作者：
DebBurman, SK
中科院分区：
文献类型：
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作者：
Brandis, KA;Holmes, IF;DebBurman, SK
尽管裂变酵母的建模突出的细胞过程的历史，它还没有被用来模拟人类神经变性相关的蛋白质错误折叠。由于α-突触核蛋白错误折叠和聚集与帕金森病（PD）有关，因此在这里，我们报告了一种裂变酵母（粟酒裂殖酵母）模型，该模型评估了α-突触核蛋白错误折叠、聚集和毒性，并将这些特性与最近在芽殖酵母中表征的特性进行比较（酿酒酵母）。野生型α-突触核蛋白和三种突变体（A30 P、A53 T和A30 P/A53 T）用硫胺素阻遏型启动子（使用启动子强度增加的载体：pNMT 81、pNMT 41和pNMT 1）表达，以直接在活细胞中测试α-突触核蛋白错误折叠和聚集的成核聚合假说。在支持的假设，野生型和A53 T α-突触核蛋白形成明显的细胞内的细胞质内含物内的裂变酵母细胞中的浓度和时间依赖性的方式，而A30 P和A30 P/A53 T仍然弥漫在整个细胞质。A53 T α-突触核蛋白比野生型α-突触核蛋白更快地形成聚集体，并且在较低的α-突触核蛋白浓度下。出乎意料的是，与芽殖酵母中不同，野生型和A53 T α-突触核蛋白不靶向裂殖酵母中的质膜，甚至在低α-突触核蛋白浓度下或作为形成聚集体的前体步骤也不靶向质膜。尽管α-突触核蛋白广泛聚集，但令人惊讶的是，它对裂变酵母无毒。未来的基因解剖可能会产生对这种毒性保护的分子见解。我们推测α-突触核蛋白的毒性可能与其膜结合能力有关。最后，S. pombe和S.酿酒酵母模型相似但不同方面的α-突触核蛋白生物学，和两种生物体揭示了α-突触核蛋白的作用，在PD发病机制。
Despite fission yeast's history of modeling salient cellular processes, it has not yet been used to model human neurodegeneration-linked protein misfolding. Because alpha-synuclein misfolding and aggregation are linked to Parkinson's disease (PD), here, we report a fission yeast (Schizosaccharomyces pombe) model that evaluates alpha-synuclein misfolding, aggregation, and toxicity and compare these properties with those recently characterized in budding yeast (Saccharomyces cerevisiae). Wild-type alpha-synuclein and three mutants (A30P, 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 a-synuclein misfolding and aggregation. In support of the hypothesis, wild-type and A53T alpha-synuclein formed prominent intracellular cytoplasmic inclusions within fission yeast cells in a concentration- and time-dependent manner, whereas A30P and A30P/A53T remained diffuse throughout the cytoplasm. A53T alpha-synuclein formed aggregates faster than wild-type a-synuclein and at a lower alpha-synuclein concentration. Unexpectedly, unlike in budding yeast, wild-type and A53T alpha-synuclein did not target to the plasma membrane in fission yeast, not even at low alpha-synuclein concentrations or as a precursor step to forming aggregates. Despite alpha-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 alpha-synuclein toxicity might be linked to its membrane binding capacity. To conclude, S. pombe and S. cerevisiae model similar yet distinct aspects of alpha-synuclein biology, and both organisms shed insight into alpha-synuclein's role in PD pathogenesis.