Hypoculoside, a sphingoid base-like compound from Acremonium disrupts the membrane integrity of yeast cells

Abstract

We have isolated Hypoculoside, a new glycosidic amino alcohol lipid from the fungus Acremonium sp. F2434 belonging to the order Hypocreales and determined its structure by 2D-NMR (Nuclear Magnetic Resonance) spectroscopy. Hypoculoside has antifungal, antibacterial and cytotoxic activities. Homozygous profiling (HOP) of hypoculoside in Saccharomyces cerevisiae (budding yeast) revealed that several mutants defective in vesicular trafficking and vacuolar protein transport are sensitive to hypoculoside. Staining of budding yeast cells with the styryl dye FM4-64 indicated that hypoculoside damaged the vacuolar structure. Furthermore, the propidium iodide (PI) uptake assay showed that hypoculoside disrupted the plasma membrane integrity of budding yeast cells. Interestingly, the glycosidic moiety of hypoculoside is required for its deleterious effect on growth, vacuoles and plasma membrane of budding yeast cells.

Chemogenomic profiling in yeast reveals antifungal mode-of-action of polyene macrolactam auroramycin

“…Auroramycin was used at 1.56 μM which caused a 50% reduction in growth in YPD-HEPES medium after 10 hours at 30 °C. Preparation of genomic DNA from yeast cells, amplification of barcode sequences by PCR and Next Generation Sequencing (NGS) of the PCR products were all performed as previously described [14].…”

Diethyl phthalate (DEP) perturbs nitrogen metabolism in Saccharomyces cerevisiae

“…Some of the 4817 mutants had a significantly high absolute logFC value. Mutants with positive logFC values in homozygous profiling assays exhibit virtually no resistance to the compound from our previous work 25 , 26 , 35 . Henceforth, we focused on DEP-sensitive mutants (i.e., mutants with logFC value < − 0.5).…”

“…Intriguingly, the relative toxicity of another phthalate diester and monoester pair is exactly the opposite. Mono(2-ethylhexyl) phthalate (MEHP) is more potent than its corresponding diester, di-(2-ethylhexyl)-phthalate (DEHP), in yeast and mammalian cells 25 , 26 . Our previous studies showed that MEHP affected plasma membrane integrity.…”

“…Chemogenomic profiling was performed using a pooled barcoded library derived from the Saccharomyces cerevisiae diploid homozygous deletion collection (Invitrogen) as described previously 25 , 26 . Pooled library of barcoded deletion strains was treated with either DMSO or 0.78 mM DEP.…”

Sophorolipid exhibits antifungal activity by ROS mediated endoplasmic reticulum stress and mitochondrial dysfunction pathways in Candida albicans

“…The Propidium Iodide (PI) staining assay was carried out as described by Alfatah et al 29 with slight modification. Briefly, C. albicans SC5314 cells of 0.5 OD were treated with different concentration of SL along with DMSO as vehicle control in 2 mL of YPD medium at 30 °C and 200 rpm for 5 h. Further, the treated cells were washed twice and resuspended in 1× PBS.…”