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dc.contributor.authorReis, Flavia Coelho Garcia dospt_BR
dc.contributor.authorBorges, Beatriz Santanapt_BR
dc.contributor.authorJozefowicz, Luísa Jennrichpt_BR
dc.contributor.authorSena, Bianca Aparecida Gimenez dept_BR
dc.contributor.authorGarcia, Ane Wichine Acostapt_BR
dc.contributor.authorMedeiros, Lia Carolina Soarespt_BR
dc.contributor.authorMartins, Sharon de Toledopt_BR
dc.contributor.authorHonorato, Leandropt_BR
dc.contributor.authorSchrank, Augustopt_BR
dc.contributor.authorVainstein, Marilene Henningpt_BR
dc.contributor.authorSilva, Lívia Kmetzsch Rosa ept_BR
dc.contributor.authorNimrichter, Leonardopt_BR
dc.contributor.authorAlves, Lysangela Ronaltept_BR
dc.contributor.authorStaats, Charley Christianpt_BR
dc.contributor.authorRodrigues, Marcio Lourençopt_BR
dc.date.accessioned2023-11-30T03:23:05Zpt_BR
dc.date.issued2019pt_BR
dc.identifier.issn2379-5042pt_BR
dc.identifier.urihttp://hdl.handle.net/10183/267818pt_BR
dc.description.abstractRegular protocols for the isolation of fungal extracellular vesicles (EVs) are time-consuming, hard to reproduce, and produce low yields. In an attempt to improve the protocols used for EV isolation, we explored a model of vesicle produc tion after growth of Cryptococcus gattii and Cryptococcus neoformans on solid media. Nanoparticle tracking analysis in combination with transmission electron microscopy revealed that C. gattii and C. neoformans produced EVs in solid media. The proper ties of cryptococcal vesicles varied according to the culture medium used and the EV-producing species. EV detection was reproduced with an acapsular mutant of C. neoformans, as well as with isolates of Candida albicans, Histoplasma capsulatum, and Saccharomyces cerevisiae. Cryptococcal EVs produced in solid media were bio logically active and contained regular vesicular components, including the major polysaccharide glucuronoxylomannan (GXM) and RNA. Since the protocol had higher yields and was much faster than the regular methods used for the isolation of fun gal EVs, we asked if it would be applicable to address fundamental questions related to cryptococcal secretion. On the basis that polysaccharide export in Cryptococcus requires highly organized membrane traffic culminating with EV release, we ana lyzed the participation of a putative scramblase (Aim25; CNBG_3981) in EV-mediated GXM export and capsule formation in C. gattii. EVs from a C. gattii aim25Δ strain dif fered from those obtained from wild-type (WT) cells in physical-chemical properties and cargo. In a model of surface coating of an acapsular cryptococcal strain with ve sicular GXM, EVs obtained from the aim25Δ mutant were more efficiently used as a source of capsular polysaccharides. Lack of the Aim25 scramblase resulted in disor ganized membranes and increased capsular dimensions. These results associate the description of a novel protocol for the isolation of fungal EVs with the identification of a previously unknown regulator of polysaccharide release.en
dc.format.mimetypeapplication/pdfpt_BR
dc.language.isoengpt_BR
dc.relation.ispartofmSphere. Washington, DC. Vol. 4, no. 2 (Mar./Apr 2019), e00080-19, 15 p.pt_BR
dc.rightsOpen Accessen
dc.subjectCryptococcus gattiipt_BR
dc.subjectCryptococcusen
dc.subjectCryptococcus neoformanspt_BR
dc.subjectExtracellular vesiclesen
dc.subjectFungien
dc.subjectVirulênciapt_BR
dc.subjectVesículas extracelularespt_BR
dc.subjectSecretionen
dc.subjectScramblaseen
dc.titleA novel protocol for the isolation of fungal extracellular vesicles reveals the participation of a putative scramblase in polysaccharide export and capsule construction in Cryptococcus gattiipt_BR
dc.typeArtigo de periódicopt_BR
dc.identifier.nrb001172911pt_BR
dc.type.originEstrangeiropt_BR


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