Identification of novel parasitophorous vacuole proteins in P. falciparum parasites using BioID
Identification of novel parasitophorous vacuole proteins in P. falciparum parasites using BioID
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DOI：
10.1016/j.ijmm.2017.07.007
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发表时间：
2018-01-01
影响因子：
4.1
通讯作者：
Spielmann, Tobias
中科院分区：
文献类型：
--
作者：
Khosh-Naucke, Melissa;Beeker, Johanna;Spielmann, Tobias
疟疾血液阶段寄生虫在红细胞内发育，它们被包含在称为寄生泡（PV）的液泡室中。该区室在寄生虫与其宿主细胞的相互作用中发挥着关键作用。然而，迄今为止，该区室的蛋白质组尚未得到全面分析。在这里，我们在最具毒性的人类疟原虫恶性疟原虫的无性血液阶段使用 BioID 来鉴定 PV 的新蛋白质。由此产生的蛋白质组包含许多已知的 PV 蛋白，并且通过 GFP 敲入先前在恶性疟原虫中未表征的命中的 10 个蛋白进行验证，揭示了 5 个新的 PV 蛋白和两个具有部分 PV 定位的蛋白。这包括附着在PV膜内表面的外围蛋白质以及锚定在突出到PV中的寄生虫质膜中的蛋白质。通过选择性的靶向基因破坏，我们为 10 个候选者中的 2 个产生了突变体。相比之下，我们无法为另外 3 种候选者选择受到干扰的寄生虫，这强烈表明它们对寄生虫的生长很重要。有趣的是，其中之一包括 UIS2 的直系同源物，这是一种先前提出调节寄生虫细胞质中蛋白质翻译的蛋白质，但这里显示它是一种必需的 PV 蛋白质。这项工作扩展了已知 PV 蛋白的数量，并为该区室的进一步功能分析提供了起点。
Malaria blood stage parasites develop within red blood cells where they are contained in a vacuolar compartment known as the parasitophorous vacuole (PV). This compartment holds a key role in the interaction of the parasite with its host cell. However, the proteome of this compartment has so far not been comprehensively analysed. Here we used BioID in asexual blood stages of the most virulent human malaria parasite Plasmodium falciparum to identify new proteins of the PV. The resulting proteome contained many of the already known PV proteins and validation by GFP-knock-in of 10 previously in P. falciparum uncharacterised hits revealed 5 new PV proteins and two with a partial PV localisation. This included proteins peripherally attached to the inner face of the PV membrane as well as proteins anchored in the parasite plasma membrane that protrude into the PV. Using selectable targeted gene disruption we generated mutants for 2 of the 10 candidates. In contrast we could not select parasites with disruptions for another 3 candidates, strongly suggesting that they are important for parasite growth. Interestingly, one of these included the orthologue of UIS2, a protein previously proposed to regulate protein translation in the parasite cytoplasm but here shown to be an essential PV protein. This work extends the number of known PV proteins and provides a starting point for further functional analyses of this compartment.