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EXP1 is required for organisation of EXP2 in the intraerythrocytic malaria parasite vacuole.
Cellular Microbiology ( IF 2.6 ) Pub Date : 2020-02-12 , DOI: 10.1111/cmi.13168
Timothy Nessel 1 , John M Beck 1 , Shima Rayatpisheh 2 , Yasaman Jami-Alahmadi 2 , James A Wohlschlegel 2 , Daniel E Goldberg 3 , Josh R Beck 1, 3
Affiliation  

Intraerythrocytic malaria parasites reside within a parasitophorous vacuole membrane (PVM) that closely overlays the parasite plasma membrane. Although the PVM is the site of several transport activities essential to parasite survival, the basis for organisation of this membrane system is unknown. Here, we performed proximity labeling at the PVM with BioID2, which highlighted a group of single-pass integral membrane proteins that constitute a major component of the PVM proteome but whose function remains unclear. We investigated EXP1, the longest known member of this group, by adapting a CRISPR/Cpf1 genome editing system to install the TetR-DOZI-aptamers system for conditional translational control. Importantly, although EXP1 was required for intraerythrocytic development, a previously reported in vitro glutathione S-transferase activity could not account for this essential EXP1 function in vivo. EXP1 knockdown was accompanied by profound changes in vacuole ultrastructure, including apparent increased separation of the PVM from the parasite plasma membrane and formation of abnormal membrane structures. Furthermore, although activity of the Plasmodium translocon of exported proteins was not impacted by depletion of EXP1, the distribution of the translocon pore-forming protein EXP2 but not the HSP101 unfoldase was substantially altered. Collectively, our results reveal a novel PVM defect that indicates a critical role for EXP1 in maintaining proper organisation of EXP2 within the PVM.

中文翻译:

EXP1是在红细胞内疟原虫空泡中组织EXP2所必需的。

红细胞内疟原虫位于寄生虫液泡膜(PVM)中,该膜紧密覆盖寄生虫的质膜。尽管PVM是寄生虫生存必不可少的几种运输活动的场所,但该膜系统的组织基础尚不清楚。在这里,我们用BioID2在PVM上进行了邻近标记,该标记突出了一组单程整合膜蛋白,这些蛋白构成了PVM蛋白质组的主要成分,但其​​功能尚不清楚。我们通过调整CRISPR / Cpf1基因组编辑系统以安装TetR-DOZI-aptamers系统进行条件翻译控制,研究了该组中最长的成员EXP1。重要的是,尽管EXP1是红细胞内发育所必需的,先前报道的体外谷胱甘肽S-转移酶活性不能解释体内这种必需的EXP1功能。EXP1击倒伴随着液泡超微结构的深刻变化,包括PVM与寄生虫质膜的明显分离和异常膜结构的形成。此外,尽管出口蛋白的疟原虫转座子的活性不受EXP1的消耗的影响,但形成转座子的孔形成蛋白EXP2的分布却不受HSP101解折叠酶的影响。总的来说,我们的结果揭示了一个新的PVM缺陷,该缺陷指示EXP1在维持EXPM在PVM中的正确组织中的关键作用。包括PVM与寄生虫质膜的明显增加分离以及异常膜结构的形成。此外,尽管出口蛋白的疟原虫转座子的活性不受EXP1的消耗的影响,但形成转座子的孔形成蛋白EXP2的分布却不受HSP101解折叠酶的影响。总的来说,我们的结果揭示了一个新的PVM缺陷,该缺陷指示EXP1在维持EXPM在PVM中的正确组织中的关键作用。包括PVM与寄生虫质膜的明显增加分离以及异常膜结构的形成。此外,尽管出口蛋白的疟原虫转座子的活性不受EXP1的消耗的影响,但形成转座子的孔形成蛋白EXP2的分布却不受HSP101解折叠酶的影响。总的来说,我们的结果揭示了一个新的PVM缺陷,该缺陷指示EXP1在维持EXPM在PVM中的正确组织中的关键作用。Translocon孔形成蛋白EXP2的分布,但没有改变HSP101解折叠酶。总的来说,我们的结果揭示了一个新的PVM缺陷,该缺陷指示EXP1在维持EXPM在PVM中的正确组织中的关键作用。Translocon孔形成蛋白EXP2的分布,但没有改变HSP101解折叠酶。总的来说,我们的结果揭示了一个新的PVM缺陷,该缺陷指示EXP1在维持EXPM在PVM中的正确组织中的关键作用。
更新日期:2020-02-12
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