RT Journal Article SR Electronic T1 The ESCRT-III machinery participates in the production of extracellular vesicles and protein export during Plasmodium falciparum infection JF bioRxiv FD Cold Spring Harbor Laboratory SP 2020.11.24.395756 DO 10.1101/2020.11.24.395756 A1 Avalos-Padilla, Yunuen A1 Georgiev, Vasil N. A1 Lantero, Elena A1 Pujals, Silvia A1 Verhoef, René A1 Borgheti-Cardoso, Livia N. A1 Albertazzi, Lorenzo A1 Dimova, Rumiana A1 Fernàndez-Busquets, Xavier YR 2020 UL http://biorxiv.org/content/early/2020/11/24/2020.11.24.395756.abstract AB Infection with Plasmodium falciparum enhances extracellular vesicles (EVs) production in parasitized red blood cells (pRBC), 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 that is activated by an alternative recruitment pathway involving the action of PfBro1 and PfVps32/PfVps60 proteins. Additionally, multivesicular bodies formation and membrane shedding, both reported mechanisms of EVs 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. In accordance, disruption of the Pfvps60 gene led to a reduction in the number of the produced EVs in the KO strain when compared with the parental 3D7 strain. Overall, our results increase the knowledge on the underlying molecular mechanisms during malaria pathogenesis and demonstrate that ESCRT-III P. falciparum proteins participate in EVs production.Competing Interest StatementThe authors have declared no competing interest.