Insights into dynamin-associated disorders through analysis of equivalent mutations in the yeast dynamin Vps1.
Insights into dynamin-associated disorders through analysis of equivalent mutations in the yeast dynamin Vps1.
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DOI：
10.15698/mic2016.04.490
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发表时间：
2016-03-22
期刊：
影响因子：
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通讯作者：
Ayscough KR
中科院分区：
文献类型：
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作者：
Moustaq L;Smaczynska-de Rooij II;Palmer SE;Marklew CJ;Ayscough KR
动力素代表了一个蛋白质超家族，已被证明在广泛的膜融合和裂变事件中发挥作用。据报道，人类经典动力素dyn-1和dyn-2发生了越来越多的突变，由这些突变引起的疾病从Charcot-Marie-Tooth疾病到癫痫脑病。发芽酵母酿酒酵母表达一种与动力蛋白相关的蛋白，该蛋白在细胞膜运输过程中起着与dyn-1和dyn-2类似的作用，在质膜上分裂内吞泡时发挥着类似的作用。动力蛋白的大部分在物种之间高度保守，这使得我们在这项研究中能够选择一些致病突变，并在Vps1中产生同等的突变。然后，我们使用细胞和生化分析对这些突变体进行了研究，以确定受这些变化影响的蛋白质的功能。具体地说，我们证明了Vps1-G397R突变(dyn-2 G358R)破坏了蛋白质的寡聚，Vps1-A447T(dyn-1 A408T)影响内吞作用的分裂阶段，而Vps1-R298L(dyn-1 R256L)影响脂结合特异性，并可能影响内吞作用的早期阶段。总体而言，我们认为酵母模型将潜在地为快速分析新的动力素突变提供一种途径，以便了解它们破坏的潜在机制。
The dynamins represent a superfamily of proteins that have been shown to function in a wide range of membrane fusion and fission events. An increasing number of mutations in the human classical dynamins, Dyn-1 and Dyn-2 has been reported, with diseases caused by these changes ranging from Charcot-Marie-Tooth disorder to epileptic encephalopathies. The budding yeast, Saccharomyces cerevisiae expresses a single dynamin-related protein that functions in membrane trafficking, and is considered to play a similar role to Dyn-1 and Dyn-2 during scission of endocytic vesicles at the plasma membrane. Large parts of the dynamin protein are highly conserved across species and this has enabled us in this study to select a number of disease causing mutations and to generate equivalent mutations in Vps1. We have then studied these mutants using both cellular and biochemical assays to ascertain functions of the protein that have been affected by the changes. Specifically, we demonstrate that the Vps1-G397R mutation (Dyn-2 G358R) disrupts protein oligomerization, Vps1-A447T (Dyn-1 A408T) affects the scission stage of endocytosis, while Vps1-R298L (Dyn-1 R256L) affects lipid binding specificity and possibly an early stage in endocytosis. Overall, we consider that the yeast model will potentially provide an avenue for rapid analysis of new dynamin mutations in order to understand the underlying mechanisms that they disrupt