The ESCRT-III machinery participates in the production of extracellular vesicles and protein export during Plasmodium falciparum infection.
The ESCRT-III machinery participates in the production of extracellular vesicles and protein export during Plasmodium falciparum infection.
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DOI：
10.1371/journal.ppat.1009455
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发表时间：
2021-04
期刊：
影响因子：
6.7
通讯作者：
中科院分区：
文献类型：
--
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恶性疟原虫感染可增加红细胞(PRBC)胞外小泡(EV)的产生，这是红细胞内无性繁殖过程中寄生虫与寄生虫交流的重要机制。转运所需的内体分选复合体(ESCRT)，特别是ESCRT-III亚复合体，参与高等真核生物EVS的形成。然而，红细胞在成熟过程中失去了大部分细胞器，包括其囊泡网络的重要减少。因此，恶性疟原虫感染红细胞产生EV的机制尚不清楚。在这里，我们证明了恶性疟原虫具有一个功能性的ESCRT-III机制，该机制由PfBro1和PfVps32/PfVps60蛋白的另一种募集途径激活。此外，利用纯化的重组蛋白在巨大单层囊泡的膜模型中重组了已报道的EV产生的两种机制--多囊泡体形成和膜脱落。此外，超分辨显微镜和斑点印迹分析证实，从pRBC培养中纯化的EV中存在PfVps32、PfVps60和PfBro1。最后，PfVps60基因的破坏导致KO株中产生的EVS数量减少，并影响了ESCRT-III的其他成分的分布。总体而言，我们的结果增加了对疟疾发病过程中潜在分子机制的了解，并表明ESCRT-III恶性疟原虫蛋白参与了EV的产生。疟疾是由疟原虫引起的一种疾病，在许多低收入国家，这种疾病仍然是主要的死亡原因，目前可用的治疗策略在控制这种疾病方面并不成功，更不用说根除了。在疟原虫入侵后观察到的一个有趣的特征是寄生的红细胞(PRBC)产生的细胞外小泡(EV)增加，这缺乏解释EV产生的小泡运输。在这里，通过结合不同的方法，我们证明了恶性疟原虫运输所需的内体分选复合体(ESCRT)机械参与了pRBC中EVS的生产。此外，我们还能够在宿主膜附近检测到ESCRT-III蛋白，并在从pRBC培养中纯化的EV中检测到ESCRT-III蛋白，这表明这些蛋白的输出及其参与EV的产生。最后，ESCRT-III相关基因Pfvps60的中断导致EV数量显著减少。总之，这些结果证实了ESCRT-III参与了EV的产生，并提供了关于恶性疟原虫蛋白输出机制的新信息，这些机制可用于在中断EV形成和贩运的基础上制定新的疟疾治疗策略。
Infection with Plasmodium falciparum enhances extracellular vesicle (EV) production in parasitized red blood cells (pRBCs), an important mechanism for parasite-to-parasite communication during the asexual intraerythrocytic life cycle. The endosomal sorting complex required for transport (ESCRT), and in particular the ESCRT-III sub-complex, participates in the formation of EVs in higher eukaryotes. However, RBCs have lost the majority of their organelles through the maturation process, including an important reduction in their vesicular network. Therefore, the mechanism of EV production in P. falciparum-infected RBCs remains to be elucidated. Here we demonstrate that P. falciparum possesses a functional ESCRT-III machinery activated by an alternative recruitment pathway involving the action of PfBro1 and PfVps32/PfVps60 proteins. Additionally, multivesicular body formation and membrane shedding, both reported mechanisms of EV production, were reconstituted in the membrane model of giant unilamellar vesicles using the purified recombinant proteins. Moreover, the presence of PfVps32, PfVps60 and PfBro1 in EVs purified from a pRBC culture was confirmed by super-resolution microscopy and dot blot assays. Finally, disruption of the PfVps60 gene led to a reduction in the number of the produced EVs in the KO strain and affected the distribution of other ESCRT-III components. Overall, our results increase the knowledge on the underlying molecular mechanisms during malaria pathogenesis and demonstrate that ESCRT-III P. falciparum proteins participate in EV production. Malaria is a disease caused by Plasmodium parasites that is still a leading cause of death in many low-income countries, and for which currently available therapeutic strategies are not succeeding in its control, let alone eradication. An interesting feature observed after Plasmodium invasion is the increase of extracellular vesicles (EVs) generated by parasitized red blood cells (pRBCs), which lack a vesicular trafficking that would explain EV production. Here, by combining different approaches, we demonstrated the participation of the endosomal sorting complex required for transport (ESCRT) machinery from Plasmodium falciparum in the production of EVs in pRBCs. Moreover, we were able to detect ESCRT-III proteins adjacent to the membrane of the host and in EVs purified from a pRBC culture, which shows the export of these proteins and their participation in EV production. Finally, the disruption of an ESCRT-III associated gene, Pfvps60, led to a significant reduction in the amount of EVs. Altogether, these results confirm ESCRT-III participation in EV production and provide novel information on the P. falciparum protein export mechanisms, which can be used for the development of new therapeutic strategies against malaria, based on the disruption of EV formation and trafficking.
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DOI：
10.1097/qai.0000000000000756
发表时间：
2015-11-01
期刊：
Journal of acquired immune deficiency syndromes (1999)
影响因子：
--
作者：
Hubert A;Subra C;Jenabian MA;Tremblay Labrecque PF;Tremblay C;Laffont B;Provost P;Routy JP;Gilbert C
通讯作者：
Gilbert C
影响因子：
3
作者：
Campos FM;Franklin BS;Teixeira-Carvalho A;Filho AL;de Paula SC;Fontes CJ;Brito CF;Carvalho LH
通讯作者：
Carvalho LH
影响因子：
5.7
作者：
Boura, Evzen;Rozycki, Bartosz;Chung, Hoi Sung;Herrick, Dawn Z.;Canagarajah, Bertram;Cafiso, David S.;Eaton, William A.;Hummer, Gerhard;Hurley, James H.
通讯作者：
Hurley, James H.
影响因子：
46.9
作者：
Ghorbal, Mehdi;Gorman, Molly;Lopez-Rubio, Jose-Juan
通讯作者：
Lopez-Rubio, Jose-Juan
影响因子：
11.4
作者：
Gill, David J.;Teo, Hsiangling;Williams, Roger L.
通讯作者：
Williams, Roger L.