Characterization of Extracellular Vesicles Produced by <named-content content-type="genus-species">Aspergillus fumigatus</named-content> Protoplasts

ABSTRACT Extracellular vesicles (EVs) are membranous compartments produced by yeast and mycelial forms of several fungal species. One of the difficulties in perceiving the role of EVs during the fungal life, and particularly in cell wall biogenesis, is caused by the presence of a thick cell wall. On...

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Autores principales:	Juliana Rizzo, Thibault Chaze, Kildare Miranda, Robert W. Roberson, Olivier Gorgette, Leonardo Nimrichter, Mariette Matondo, Jean-Paul Latgé, Anne Beauvais, Marcio L. Rodrigues
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	Aspergillus fumigatus conidia protoplasts extracellular vesicles Aspergillus Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/72394ddcc4614f71b2f3d1f343120f61
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spelling	oai:doaj.org-article:72394ddcc4614f71b2f3d1f343120f612021-11-15T15:30:51ZCharacterization of Extracellular Vesicles Produced by <named-content content-type="genus-species">Aspergillus fumigatus</named-content> Protoplasts10.1128/mSphere.00476-202379-5042https://doaj.org/article/72394ddcc4614f71b2f3d1f343120f612020-08-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00476-20https://doaj.org/toc/2379-5042ABSTRACT Extracellular vesicles (EVs) are membranous compartments produced by yeast and mycelial forms of several fungal species. One of the difficulties in perceiving the role of EVs during the fungal life, and particularly in cell wall biogenesis, is caused by the presence of a thick cell wall. One alternative to have better access to these vesicles is to use protoplasts. This approach has been investigated here with Aspergillus fumigatus, one of the most common opportunistic fungal pathogens worldwide. Analysis of regenerating protoplasts by scanning electron microscopy and fluorescence microscopy indicated the occurrence of outer membrane projections in association with surface components and the release of particles with properties resembling those of fungal EVs. EVs in culture supernatants were characterized by transmission electron microscopy and nanoparticle tracking analysis. Proteomic and glycome analysis of EVs revealed the presence of a complex array of enzymes related to lipid/sugar metabolism, pathogenic processes, and cell wall biosynthesis. Our data indicate that (i) EV production is a common feature of different morphological stages of this major fungal pathogen and (ii) protoplastic EVs are promising tools for undertaking studies of vesicle functions in fungal cells. IMPORTANCE Fungal cells use extracellular vesicles (EVs) to export biologically active molecules to the extracellular space. In this study, we used protoplasts of Aspergillus fumigatus, a major fungal pathogen, as a model to evaluate the role of EV production in cell wall biogenesis. Our results demonstrated that wall-less A. fumigatus exports plasma membrane-derived EVs containing a complex combination of proteins and glycans. Our report is the first to characterize fungal EVs in the absence of a cell wall. Our results suggest that protoplasts represent a promising model for functional studies of fungal vesicles.Juliana RizzoThibault ChazeKildare MirandaRobert W. RobersonOlivier GorgetteLeonardo NimrichterMariette MatondoJean-Paul LatgéAnne BeauvaisMarcio L. RodriguesAmerican Society for MicrobiologyarticleAspergillus fumigatusconidiaprotoplastsextracellular vesiclesAspergillusMicrobiologyQR1-502ENmSphere, Vol 5, Iss 4 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Aspergillus fumigatus conidia protoplasts extracellular vesicles Aspergillus Microbiology QR1-502
spellingShingle	Aspergillus fumigatus conidia protoplasts extracellular vesicles Aspergillus Microbiology QR1-502 Juliana Rizzo Thibault Chaze Kildare Miranda Robert W. Roberson Olivier Gorgette Leonardo Nimrichter Mariette Matondo Jean-Paul Latgé Anne Beauvais Marcio L. Rodrigues Characterization of Extracellular Vesicles Produced by <named-content content-type="genus-species">Aspergillus fumigatus</named-content> Protoplasts
description	ABSTRACT Extracellular vesicles (EVs) are membranous compartments produced by yeast and mycelial forms of several fungal species. One of the difficulties in perceiving the role of EVs during the fungal life, and particularly in cell wall biogenesis, is caused by the presence of a thick cell wall. One alternative to have better access to these vesicles is to use protoplasts. This approach has been investigated here with Aspergillus fumigatus, one of the most common opportunistic fungal pathogens worldwide. Analysis of regenerating protoplasts by scanning electron microscopy and fluorescence microscopy indicated the occurrence of outer membrane projections in association with surface components and the release of particles with properties resembling those of fungal EVs. EVs in culture supernatants were characterized by transmission electron microscopy and nanoparticle tracking analysis. Proteomic and glycome analysis of EVs revealed the presence of a complex array of enzymes related to lipid/sugar metabolism, pathogenic processes, and cell wall biosynthesis. Our data indicate that (i) EV production is a common feature of different morphological stages of this major fungal pathogen and (ii) protoplastic EVs are promising tools for undertaking studies of vesicle functions in fungal cells. IMPORTANCE Fungal cells use extracellular vesicles (EVs) to export biologically active molecules to the extracellular space. In this study, we used protoplasts of Aspergillus fumigatus, a major fungal pathogen, as a model to evaluate the role of EV production in cell wall biogenesis. Our results demonstrated that wall-less A. fumigatus exports plasma membrane-derived EVs containing a complex combination of proteins and glycans. Our report is the first to characterize fungal EVs in the absence of a cell wall. Our results suggest that protoplasts represent a promising model for functional studies of fungal vesicles.
format	article
author	Juliana Rizzo Thibault Chaze Kildare Miranda Robert W. Roberson Olivier Gorgette Leonardo Nimrichter Mariette Matondo Jean-Paul Latgé Anne Beauvais Marcio L. Rodrigues
author_facet	Juliana Rizzo Thibault Chaze Kildare Miranda Robert W. Roberson Olivier Gorgette Leonardo Nimrichter Mariette Matondo Jean-Paul Latgé Anne Beauvais Marcio L. Rodrigues
author_sort	Juliana Rizzo
title	Characterization of Extracellular Vesicles Produced by <named-content content-type="genus-species">Aspergillus fumigatus</named-content> Protoplasts
title_short	Characterization of Extracellular Vesicles Produced by <named-content content-type="genus-species">Aspergillus fumigatus</named-content> Protoplasts
title_full	Characterization of Extracellular Vesicles Produced by <named-content content-type="genus-species">Aspergillus fumigatus</named-content> Protoplasts
title_fullStr	Characterization of Extracellular Vesicles Produced by <named-content content-type="genus-species">Aspergillus fumigatus</named-content> Protoplasts
title_full_unstemmed	Characterization of Extracellular Vesicles Produced by <named-content content-type="genus-species">Aspergillus fumigatus</named-content> Protoplasts
title_sort	characterization of extracellular vesicles produced by <named-content content-type="genus-species">aspergillus fumigatus</named-content> protoplasts
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/72394ddcc4614f71b2f3d1f343120f61
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Characterization of Extracellular Vesicles Produced by <named-content content-type="genus-species">Aspergillus fumigatus</named-content> Protoplasts

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