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The\n                    <jats:underline>e<\/jats:underline>\n                    ndosomal\n                    <jats:underline>s<\/jats:underline>\n                    orting\n                    <jats:underline>c<\/jats:underline>\n                    omplex\n                    <jats:underline>r<\/jats:underline>\n                    equired for\n                    <jats:underline>t<\/jats:underline>\n                    ransport (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\n                    <jats:italic>P<\/jats:italic>\n                    .\n                    <jats:italic>falciparum-<\/jats:italic>\n                    infected RBCs remains to be elucidated. Here we demonstrate that\n                    <jats:italic>P<\/jats:italic>\n                    .\n                    <jats:italic>falciparum<\/jats:italic>\n                    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\n                    <jats:italic>PfVps60<\/jats:italic>\n                    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\n                    <jats:italic>P<\/jats:italic>\n                    .\n                    <jats:italic>falciparum<\/jats:italic>\n                    proteins participate in EV production.\n                  <\/jats:p>","DOI":"10.1371\/journal.ppat.1009455","type":"journal-article","created":{"date-parts":[[2021,4,2]],"date-time":"2021-04-02T13:34:00Z","timestamp":1617370440000},"page":"e1009455","update-policy":"https:\/\/doi.org\/10.1371\/journal.ppat.corrections_policy","source":"Crossref","is-referenced-by-count":50,"title":["The ESCRT-III machinery participates in the production of extracellular vesicles and protein export during Plasmodium falciparum infection"],"prefix":"10.1371","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4074-5624","authenticated-orcid":true,"given":"Yunuen","family":"Avalos-Padilla","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vasil N.","family":"Georgiev","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0663-4683","authenticated-orcid":true,"given":"Elena","family":"Lantero","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1776-4296","authenticated-orcid":true,"given":"Silvia","family":"Pujals","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ren\u00e9","family":"Verhoef","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9655-8214","authenticated-orcid":true,"given":"Livia","family":"N. Borgheti-Cardoso","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lorenzo","family":"Albertazzi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3872-8502","authenticated-orcid":true,"given":"Rumiana","family":"Dimova","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4622-9631","authenticated-orcid":true,"given":"Xavier","family":"Fern\u00e0ndez-Busquets","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2021,4,2]]},"reference":[{"key":"ppat.1009455.ref001","author":"WHO","year":"2019"},{"issue":"1","key":"ppat.1009455.ref002","first-page":"6","article-title":"Parasite-Derived Plasma Microparticles Contribute Significantly to Malaria Infection-Induced Inflammation through Potent Macrophage Stimulation.","author":"KN Couper","year":"2010","journal-title":"Plos Pathogens"},{"issue":"21","key":"ppat.1009455.ref003","first-page":"2542","article-title":"Circulating endothelial microparticles in Malawian children with severe falciparum malaria complicated with coma.","volume":"291","author":"V Combes","year":"2004","journal-title":"Jama-Journal of the American Medical Association"},{"issue":"10","key":"ppat.1009455.ref004","article-title":"Elevated Cell-Specific Microparticles Are a Biological Marker for Cerebral Dysfunctions in Human Severe Malaria","volume":"5","author":"JBP Mfonkeu","year":"2010","journal-title":"Plos One"},{"key":"ppat.1009455.ref005","first-page":"9","article-title":"Augmented plasma microparticles during acute Plasmodium vivax infection","author":"FMF Campos","year":"2010","journal-title":"Malaria Journal"},{"issue":"5315","key":"ppat.1009455.ref006","doi-asserted-by":"crossref","first-page":"1122","DOI":"10.1126\/science.276.5315.1122","article-title":"A membrane network for nutrient import in red cells infected with the malaria parasite","volume":"276","author":"SA Lauer","year":"1997","journal-title":"Science"},{"issue":"1","key":"ppat.1009455.ref007","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/s11060-013-1084-8","article-title":"Biogenesis of extracellular vesicles (EV): exosomes, microvesicles, retrovirus-like vesicles, and apoptotic bodies","volume":"113","author":"JC Akers","year":"2013","journal-title":"Journal of Neuro-Oncology"},{"issue":"10","key":"ppat.1009455.ref008","doi-asserted-by":"crossref","DOI":"10.1101\/cshperspect.a016048","article-title":"Missing Pieces of an Ancient Puzzle: Evolution of the Eukaryotic Membrane-Trafficking System.","volume":"6","author":"A Schlacht","year":"2014","journal-title":"Cold Spring Harbor Perspectives in Biology"},{"issue":"4","key":"ppat.1009455.ref009","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1038\/nrm.2017.125","article-title":"\u2019Angelo G, Raposo G. 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