Ergosterol distribution controls surface structure formation and fungal pathogenicity

Choy, Hau Lam; Gaylord, Elizabeth A; Doering, Tamara L.

doi:10.1101/2023.02.17.528979

Cited by 7 publications

(11 citation statements)

References 70 publications

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“…Cells lacking this gene (ysp2Δ) accumulate ergosterol at the plasma membrane, leading to dramatic deformations of the membrane and cell wall. These mutant cells are also more sensitive than wild-type to the steroltargeting antifungal compound AmB and other membrane stressors 52 . To measure the response of this mutant to BRI and CAS, we performed MIC assays, using YNB medium because ysp2Δ cells grow extremely poorly in RPMI 52 .…”

Section: Resultsmentioning

confidence: 99%

“…These mutant cells are also more sensitive than wild-type to the steroltargeting antifungal compound AmB and other membrane stressors 52 . To measure the response of this mutant to BRI and CAS, we performed MIC assays, using YNB medium because ysp2Δ cells grow extremely poorly in RPMI 52 . We found that the MIC for each compound was higher for ysp2Δ than for the corresponding wild-type, KN99a: 32 versus 8 µg/mL for CAS (Figure 2c) and 2.5 versus 1.25 µM for BRI (Figure 2d).…”

Section: Resultsmentioning

confidence: 99%

“…The YSP 2 gene of C. neoformans encodes a retrograde sterol transporter 52 . Cells lacking this gene ( ysp2 Δ) accumulate ergosterol at the plasma membrane, leading to dramatic deformations of the membrane and cell wall.…”

Section: Resultsmentioning

confidence: 99%

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Brilacidin, a novel antifungal agent againstCryptococcus neoformans

Diehl,

Pinzan,

de Castro

et al. 2024

Preprint

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Cryptococcus neoformanscauses cryptococcosis, one of the most prevalent fungal diseases, generally characterized by meningitis. There is a limited and not very effective number of drugs available to combat this disease. In this manuscript, we show the host defense peptide mimetic brilacidin (BRI) as a promising antifungal drug againstC. neoformans. BRI is able to affect the organization of the cell membrane, increasing fungal cell permeability. We also investigated the effects of BRI against the model systemSaccharomyces cerevisiaeby analyzing libraries of mutants grown in the presence of BRI. InS. cerevisiae, BRI also affects the cell membrane organization, but in addition the cell wall integrity pathway and calcium metabolism. In vivo experiments show BRI significantly reducesC. neoformanssurvival inside macrophages and partially clearsC. neoformanslung infection in an immunocompetent murine model of invasive pulmonary cryptococcosis. We also observed that BRI interacts with caspofungin (CAS) and amphotericin (AmB), potentiating their mechanism of action againstC. neoformans. BRI+CAS affects endocytic movement, calcineurin, and mitogen activated protein kinases. Our results indicate that BRI is a novel antifungal drug against cryptococcosis.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Brilacidin, a novel antifungal agent againstCryptococcus neoformans

Diehl,

Pinzan,

de Castro

et al. 2024

Preprint

Self Cite

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“…For quantification of morphology, a minimum of 94 cells from randomly-chosen slide coordinates were examined. For electron microscopy, cells were grown overnight in YPD or induced for 24 hours in DMEM before processing as in (48), except that the first incubation in 0.1 M sodium cacodylate buffer was performed for 2 h at RT. Supplemental Table S3.…”

Section: Imagingmentioning

confidence: 99%

Sac1 links phosphoinositide turnover to cryptococcal virulence

Gaylord,

Choy,

Chen

et al. 2024

Preprint

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Cryptococcus neoformansis an environmentally-acquired fungal pathogen that causes over 140,000 deaths per year. Cryptococcal infection occurs when infectious particles are deposited into the lung, where they encounter host phagocytic cells.C. neoformansmay be engulfed by these phagocytes, an important step of infection that leads to outcomes ranging from termination of infection to cryptococcal dissemination. To study this critical process, we screened approximately 4,700 cryptococcal gene deletion mutants for altered uptake, using primary mouse and human phagocytic cells. Among the hits of this screen, we identified 93 mutants with perturbed uptake in both systems, as well as others with differences in uptake by only one cell type. We further screened the hits for changes in thickness of the capsule, a protective polysaccharide layer around the cell which is an important cryptococcal virulence factor. This second screen yielded 131 mutants, including one lacking the phosphatidylinositol-4-phosphate phosphatase Sac1. In this work, we implicate Sac1 in both host cell uptake and capsule production. We found thatsac1mutants exhibit lipid trafficking defects, reductions in secretory system function, and changes in capsule size and composition. Many of these changes occur specifically in tissue culture media, highlighting the role of Sac1 phosphatase activity in responding to the stress of host-like conditions. Overall, these findings show how genome-scale screening can identify cellular factors that contribute to our understanding of cryptococcal biology and demonstrate the role of Sac1 in determining fungal virulence.

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“…In addition to thermotolerance, other aspects of the cactophilic lifestyle might be preadaptations for human pathogenicity. For instance, the sterol composition of the cell envelope has been implicated in fungal virulence 94,[116][117][118] . Mutations in genes involved in the ergosterol biosynthetic pathway are associated with antifungal resistance [118][119][120][121] , making it a major target of antifungal drugs 122 .…”

Section: Cactophily As a Launching Pad For The Emergence Of Opportuni...mentioning

confidence: 99%

Diverse signatures of convergent evolution in cacti-associated yeasts

Gonçalves,

Harrison,

Steenwyk

et al. 2023

Preprint

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Many distantly related organisms have convergently evolved traits and lifestyles that enable them to live in similar ecological environments. However, the extent of phenotypic convergence evolving through the same or distinct genetic trajectories remains an open question. Here, we leverage a comprehensive dataset of genomic and phenotypic data from 1,049 yeast species in the subphylum Saccharomycotina (Kingdom Fungi, Phylum Ascomycota) to explore signatures of convergent evolution in cactophilic yeasts, ecological specialists associated with cacti. We inferred that the ecological association of yeasts with cacti arose independently ~17 times. Using machine-learning, we further found that cactophily can be predicted with 76% accuracy from functional genomic and phenotypic data. The most informative feature for predicting cactophily was thermotolerance, which is likely associated with duplication and altered evolutionary rates of genes impacting the cell envelope in several cactophilic lineages. We also identified horizontal gene transfer and duplication events of plant cell wall-degrading enzymes in distantly related cactophilic clades, suggesting that putatively adaptive traits evolved through disparate molecular mechanisms. Remarkably, multiple cactophilic lineages and their close relatives are emerging human opportunistic pathogens, suggesting that the cactophilic lifestyle - and perhaps more generally lifestyles favoring thermotolerance - may preadapt yeasts to cause human disease. This work underscores the potential of a multifaceted approach involving high throughput genomic and phenotypic data to shed light onto ecological adaptation and highlights how convergent evolution to wild environments could facilitate the transition to human pathogenicity.

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Ergosterol distribution controls surface structure formation and fungal pathogenicity

Cited by 7 publications

References 70 publications

Brilacidin, a novel antifungal agent againstCryptococcus neoformans

Brilacidin, a novel antifungal agent againstCryptococcus neoformans

Sac1 links phosphoinositide turnover to cryptococcal virulence

Diverse signatures of convergent evolution in cacti-associated yeasts

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