Characterization of Alternaria infectoria extracellular vesicles

Silva, Branca M.A.; Prados-Rosales, Rafael; Espadas-Moreno, Javier; Wolf, Julie M.; Luque-García, José L.; Gonçalves, Teresa; Casadevall, Arturo

doi:10.1093/mmy/myt003

Cited by 77 publications

(80 citation statements)

References 42 publications

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“…Like for those previously described in C. neoformans and other fungal species, we found a large functional distribution of identified hits (23)(24)(25)(26)(27). The distribution carried through all the size-enriched populations, suggesting that any loading mechanism is size independent.…”

Section: Discussionsupporting

confidence: 84%

Interaction of Cryptococcus neoformans Extracellular Vesicles with the Cell Wall

et al. 2014

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bCryptococcus neoformans produces extracellular vesicles containing a variety of cargo, including virulence factors. To become extracellular, these vesicles not only must be released from the plasma membrane but also must pass through the dense matrix of the cell wall. The greatest unknown in the area of fungal vesicles is the mechanism by which these vesicles are released to the extracellular space given the presence of the fungal cell wall. Here we used electron microscopy techniques to image the interactions of vesicles with the cell wall. Our goal was to define the ultrastructural morphology of the process to gain insights into the mechanisms involved. We describe single and multiple vesicle-leaving events, which we hypothesized were due to plasma membrane and multivesicular body vesicle origins, respectively. We further utilized melanized cells to "trap" vesicles and visualize those passing through the cell wall. Vesicle size differed depending on whether vesicles left the cytoplasm in single versus multiple release events. Furthermore, we analyzed different vesicle populations for vesicle dimensions and protein composition. Proteomic analysis tripled the number of proteins known to be associated with vesicles. Despite separation of vesicles into batches differing in size, we did not identify major differences in protein composition. In summary, our results indicate that vesicles are generated by more than one mechanism, that vesicles exit the cell by traversing the cell wall, and that vesicle populations exist as a continuum with regard to size and protein composition.

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Section: Discussionsupporting

confidence: 84%

Interaction of Cryptococcus neoformans Extracellular Vesicles with the Cell Wall

et al. 2014

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“…Fungal EVs contain a complex combination of macromolecules, including many proteins, neutral lipids, glycans and pigments347910141518. This complex molecular mixture, including many cytoplasmic components, is consistent with the proposed origin of fungal EVs as cytoplasmic subtractions19.…”

supporting

confidence: 68%

“…Fungal cells are encased within a dense cell wall, so mechanisms are required for the transport of molecules across the wall for extracellular release in these organisms. In recent years, a number of studies have shown that the trafficking of molecules across the fungal cell wall requires extracellular vesicles (EVs)3456789101112131415161718.…”

mentioning

confidence: 99%

Extracellular vesicle-mediated export of fungal RNA

Silva

Puccia

Rodrigues

et al. 2015

Sci Rep

179

213

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Extracellular vesicles (EVs) play an important role in the biology of various organisms, including fungi, in which they are required for the trafficking of molecules across the cell wall. Fungal EVs contain a complex combination of macromolecules, including proteins, lipids and glycans. In this work, we aimed to describe and characterize RNA in EV preparations from the human pathogens Cryptococcus neoformans, Paracoccidiodes brasiliensis and Candida albicans, and from the model yeast Saccharomyces cerevisiae. The EV RNA content consisted mostly of molecules less than 250 nt long and the reads obtained aligned with intergenic and intronic regions or specific positions within the mRNA. We identified 114 ncRNAs, among them, six small nucleolar (snoRNA), two small nuclear (snRNA), two ribosomal (rRNA) and one transfer (tRNA) common to all the species considered, together with 20 sequences with features consistent with miRNAs. We also observed some copurified mRNAs, as suggested by reads covering entire transcripts, including those involved in vesicle-mediated transport and metabolic pathways. We characterized for the first time RNA molecules present in EVs produced by fungi. Our results suggest that RNA-containing vesicles may be determinant for various biological processes, including cell communication and pathogenesis.

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“…The first filamentous fungus from which EVs were isolated was Alternaria infectoria , a ubiquitous environmental fungus that spoils food crops and causes pulmonary infections in humans. Morphologically, the A. infectoria EVs resemble EVs from yeast species and are enriched with proteins that function in polysaccharide metabolism and DNA repair …”

Section: Filamentous Fungimentioning

confidence: 99%

Fungal Extracellular Vesicles with a Focus on Proteomic Analysis

2019

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Extracellular vesicles (EVs) perform crucial functions in cell–cell communication. The packaging of biomolecules into membrane‐enveloped vesicles prior to release into the extracellular environment provides a mechanism for coordinated delivery of multiple signals at high concentrations that is not achievable by classical secretion alone. Most of the understanding of the biosynthesis, composition, and function of EVs comes from mammalian systems. Investigation of fungal EVs, particularly those released by pathogenic yeast species, has revealed diverse cargo including proteins, lipids, nucleic acids, carbohydrates, and small molecules. Fungal EVs are proposed to function in a variety of biological processes including virulence and cell wall homeostasis with a focus on host–pathogen interactions. EVs also carry signals between fungal cells allowing for a coordinated attack on a host during infection. Research on fungal EVs in still in its infancy. Here a review of the literature thus far with a focus on proteomic analysis is provided with respect to techniques, results, and prospects.

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Characterization of Alternaria infectoria extracellular vesicles

Cited by 77 publications

References 42 publications

Interaction of Cryptococcus neoformans Extracellular Vesicles with the Cell Wall

Interaction of Cryptococcus neoformans Extracellular Vesicles with the Cell Wall

Extracellular vesicle-mediated export of fungal RNA

Fungal Extracellular Vesicles with a Focus on Proteomic Analysis

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