The impact of proteomics on the understanding of functions and biogenesis of fungal extracellular vesicles

Rodrigues, Márcio L.; Nakayasu, Ernesto; Almeida, Igor C.; Nimrichter, Leonardo

doi:10.1016/j.jprot.2013.04.001

Cited by 102 publications

(98 citation statements)

References 67 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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“…We identified between 179 and 278 unique proteins per strain. As with previous extracellular vesicle proteomics analyses, the proteins identified were associated with a wide variety of cellular functions and localizations (10,13,15,21,22). The 179 proteins identified in the wild-type EVs were in line with previously described C. albicans proteins (11).…”

Section: Resultssupporting

confidence: 78%

“…Recently, C. albicans EVs were found to carry several of these virulence factors and act as immunostimulants for murine immune cells (11). In addition, the membrane lipid bilayer of EVs contains fungal sterols and glucosylceramides (10,12,13). EVs from C. albicans stimulate immune responses from macrophage and dendritic cells and can contribute to a protective host effect in a waxworm model of infection (11).…”

mentioning

confidence: 99%

Lipid Biosynthetic Genes Affect Candida albicans Extracellular Vesicle Morphology, Cargo, and Immunostimulatory Properties

et al. 2015

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dMicrobial secretion is integral for regulating cell homeostasis as well as releasing virulence factors during infection. The genes encoding phosphatidylserine synthase (CHO1) and phosphatidylserine decarboxylase (PSD1 and PSD2) are Candida albicans genes involved in phospholipid biosynthesis, and mutations in these genes affect mitochondrial function, cell wall thickness, and virulence in mice. We tested the roles of these genes in several agar-based secretion assays and observed that the cho1⌬/⌬ and psd1⌬/⌬ psd2⌬/⌬ strains manifested less protease and phospholipase activity. Since extracellular vesicles (EVs) are surrounded by a lipid membrane, we investigated the effects of these mutations on EV structure, composition, and biological activity. The cho1⌬/⌬ mutant releases EVs comparable in size to wild-type EVs, but EVs from the psd1⌬/⌬ psd2⌬/⌬ strain are much larger than those from the wild type, including a population of >100-nm EVs not observed in the EVs from the wild type. Proteomic analysis revealed that EVs from both mutants had a significantly different protein cargo than that of EVs from the wild type. EVs were tested for their ability to activate NF-B in bone marrow-derived macrophage cells. While wild-type and psd1⌬/⌬ psd2⌬/⌬ mutant-derived EVs activated NF-B, the cho1⌬/⌬ mutant-derived EV did not. These studies indicate that the presence and absence of these C. albicans genes have qualitative and quantitative effects on EV size, composition, and immunostimulatory phenotypes that highlight a complex interplay between lipid metabolism and vesicle production.

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“…Several biologically relevant aspects of extracellular membrane vesicle production, both in mammalian cells and in microbes, have been highlighted in recent years. Rodrigues and colleagues [33] stress that most of the fungi reported to produce vesicles are human pathogens. Although Alternaria spp.…”

Section: Discussionmentioning

confidence: 99%

Characterization of Alternaria infectoria extracellular vesicles

Silva¹,

Prados-Rosales²,

Espadas-Moreno³

et al. 2013

Medical Mycology

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Many fungi use membrane vesicles to transport complex molecules across their cell walls. Like mammalian exosomes, fungal vesicles contain lipids, proteins, and polysaccharides, many of which are associated with virulence. Here we identify and characterize extracellular vesicles (EVs) in Alternaria infectoria, a ubiquitous, environmental filamentous fungus that is also an opportunistic human pathogen. Examination of the A. infectoria EVs revealed a morphology similar to that of vesicles described in other fungal species. Of note, proteomic analysis detected a reduced number of vesicle-associated proteins. There were two prevalent categories among the 20 identified proteins, including the polysaccharide metabolism group, probably related to plant host invasion or biosynthesis/degradation of cell wall components, and the nuclear proteins, especially DNA repair enzymes. We also found enzymes related to pigment synthesis, adhesion to the host cell, and trafficking of vesicles/organelles/molecules. This is the first time EV secretions have been identified in a filamentous fungus. We believe that these vesicles might have a role in virulence.

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The impact of proteomics on the understanding of functions and biogenesis of fungal extracellular vesicles

Cited by 102 publications

References 67 publications

Interaction of Cryptococcus neoformans Extracellular Vesicles with the Cell Wall

Interaction of Cryptococcus neoformans Extracellular Vesicles with the Cell Wall

Lipid Biosynthetic Genes Affect Candida albicans Extracellular Vesicle Morphology, Cargo, and Immunostimulatory Properties

Characterization of Alternaria infectoria extracellular vesicles

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