Vesicular mechanisms of traffic of fungal molecules to the extracellular space

Rodrigues, Márcio L.; Franzen, Anderson J.; Nimrichter, Leonardo; Miranda, Kildare

doi:10.1016/j.mib.2013.04.002

Cited by 78 publications

(78 citation statements)

References 52 publications

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“…C. neoformans vesicles have previously been morphologically characterized (15). Our images suggest that the approximately 40% observed to be electron dense may stem directly from the plasma membrane, either by pinching off or by the recently described "inverted macropinocytosis" (8), while the rest may represent vesicles from other origins.…”

Section: Discussionmentioning

confidence: 50%

“…This includes work in the model yeast Saccharomyces cerevisiae, where single deletion of genes involved with Golgi-to-plasma membrane transport or multivesicular body (MVB) formation decreased but did not halt extracellular vesicle production (7). Electron tomography has also revealed C. neoformans extracellular vesicles forming from the plasma membrane, though from membrane invagination and subsequent scission rather than the outward budding associated with bacterial outer membrane vesicles (8). Higher eukaryotes, such as Caenorhabditis elegans and Drosophila melanogaster, utilize both plasma membrane and MVBs as membrane origin sources (9,10), suggesting that eukaryotic microbes may also rely on redundant mechanisms for extracellular vesicle formation.…”

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confidence: 99%

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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: Discussionmentioning

confidence: 50%

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confidence: 99%

Interaction of Cryptococcus neoformans Extracellular Vesicles with the Cell Wall

et al. 2014

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“…We note that although no single gene has been associated with a null phenotype for vesicle production in either bacteria or fungi, several genes have now been shown to modify aspects of EV composition and structure (35,36). In this study, we report that genes involved in phospholipid synthesis can affect a variety of secretion phenotypes based not only on cell wall integrity but also on release of EVs carrying microbial cargo.…”

Section: Discussionmentioning

confidence: 79%

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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“…These findings support the growing notion that most exported proteins use alternative means of transportation, which in turn would allow for numerous cytoplasmic proteins to localize out of the plasma membrane, both in the cell wall and extracellular environment. These concepts have recently been summarized by Rodrigues et al [60], who also described a new mechanism of vesicle formation involving cell membrane invagination followed by vesicle formation in the periplasmic space of the yeast cell wall. The process involves cytoplasmic subtractions that can help explain the presence of cytoplasmic proteins on the cell wall.…”

Section: Discussionmentioning

confidence: 99%

Proteome of cell wall-extracts from pathogenic Paracoccidioides brasiliensis: Comparison among morphological phases, isolates, and reported fungal extracellular vesicle proteins

Longo

Cunha

Sobreira

et al. 2014

EuPA Open Proteomics

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Cell wall proteins Yeast Mycelium DTT Extracellular vesicles a b s t r a c tWe identified non-covalently linked cell wall proteins from Paracoccidioides brasiliensis yeasts and mycelia, with focus on the yeast pathogenic phase, and correlated them with reported fungal extracellular vesicle proteins. We studied isolates Pb3 and Pb18, which evoke distinct patterns of experimental paracoccidioidomycosis and represent two phylogenetic groups.Proteins were extracted mildly with dithiothreitol, trypsinized, and peptides analyzed by liquid chromatography coupled to high-resolution mass spectrometry. Among 132 yeastexclusive sequences, 92 were Pb18-exclusive. About 80% of total proteins were classified as secretory, mostly showing non-conventional signals. Extracellular vesicular transportation could be involved, since 60% had orthologs reported in fungal extracellular vesicles.

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Vesicular mechanisms of traffic of fungal molecules to the extracellular space

Cited by 78 publications

References 52 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

Proteome of cell wall-extracts from pathogenic Paracoccidioides brasiliensis: Comparison among morphological phases, isolates, and reported fungal extracellular vesicle proteins

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