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Heterologous expression of AtClo1, a plant oil body protein, induces lipid accumulation in yeast

机译:植物油体蛋白AtClo1的异源表达诱导酵母中脂质的积累

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Proteomic approaches on lipid bodies have led to the identification of proteins associated with this compartment, showing that, rather than the inert fat depot, lipid droplets appear as complex dynamic organelles with roles in metabolism control and cell signaling. We focused our investigations on caleosin [Arabidopsis thaliana caleosin 1 (AtClo1)], a minor protein of the Arabidopsis thaliana seed lipid body. AtClo1 shares an original triblock structure, which confers to the protein the capacity to insert at the lipid body surface. In addition, AtClo1 possesses a calcium-binding domain. The study of plants deficient in caleosin revealed its involvement in storage lipid degradation during seed germination. Using Saccharomyces cerevisiae as a heterologous expression system, we investigated the potential role of AtClo1 in lipid body biogenesis and filling. The green fluorescent protein-tagged protein was correctly targeted to lipid bodies. We observed an increase in the number and size of lipid bodies. Moreover, transformed yeasts accumulated more fatty acids (+46.6%). We confirmed that this excess of fatty acids was due to overaccumulation of lipid body neutral lipids, triacylglycerols and steryl esters. We showed that the original intrinsic properties of AtClo1 protein were sufficient to generate a functional lipid body membrane and to promote overaccumulation of storage lipids in yeast oil bodies.
机译:脂质体的蛋白质组学方法已导致鉴定了与该区室相关的蛋白质,这表明,脂滴(而不是惰性脂肪库)表现为复杂的动态细胞器,在代谢控制和细胞信号传导中起作用。我们将研究重点放在拟南芥种子脂质体的次要蛋白质caleosin [Arabidopsis thaliana caleosin 1(AtClo1)]上。 AtClo1共享原始的三嵌段结构,从而赋予蛋白质在脂质体表面插入的能力。另外,AtClo1具有钙结合结构域。缺少钙黄绿素的植物的研究表明其参与种子发芽过程中储藏脂质的降解。使用酿酒酵母作为异源表达系统,我们调查了AtClo1在脂质体的生物发生和填充中的潜在作用。带有绿色荧光蛋白标签的蛋白正确地靶向脂质体。我们观察到脂质体的数量和大小增加。此外,转化的酵母积累了更多的脂肪酸(+ 46.6%)。我们证实,脂肪酸的过量是由于脂质体中性脂质,三酰基甘油和甾醇酯的过度积累。我们表明,AtClo1蛋白的原始内在特性足以产生功能性脂质体膜并促进酵母油体中脂质的过度积累。

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