Jacko J, Morvová M, Hervay N Tóth, Eliaš D, Gbelská Y, Waczulíková I, Gášková D, Balážová M, Šikurová L
Department of Nuclear Physics and Biophysics, Faculty of Mathematics, Physics and Informatics, Comenius University Bratislava, Bratislava, Slovakia.
Department of Microbiology and Virology, Faculty of Natural Sciences, Comenius University Bratislava, Bratislava, Slovakia.
Cell Biochem Biophys. 2025 Jun;83(2):1909-1918. doi: 10.1007/s12013-024-01599-w. Epub 2024 Oct 31.
The ERG6 gene is crucial for the biosynthesis of ergosterol, a key component of yeast cell membranes. Our study examines the impact of ERG6 gene deletion on the membrane composition and physicochemical properties of the pathogenic yeast Candida glabrata. Specifically, we investigated changes in selected sterol content, phospholipid composition, transmembrane potential, and PDR16 gene activity. Sterol levels were measured using high-performance liquid chromatography, the phospholipid profile was analysed via thin-layer chromatography, transmembrane potential was assessed with fluorescence spectroscopy, and gene expression levels were determined by quantitative PCR. Our findings revealed a depletion of ergosterol, increased zymosterol and eburicol content, an increased phosphatidylcholine and a reduced phosphatidylethanolamine content in the Δerg6 strain compared to the wt. Additionally, the Δerg6 strain exhibited membrane hyperpolarization without changes in PDR16 expression. Furthermore, the Δerg6 strain showed increased sensitivity to the antifungals myriocin and aureobasidine A. These results suggest that ERG6 gene deletion leads to significant alterations in membrane composition and may activates an alternative ergosterol synthesis pathway in the C. glabrata Δerg6 deletion mutant.
ERG6基因对于麦角固醇的生物合成至关重要,麦角固醇是酵母细胞膜的关键成分。我们的研究考察了ERG6基因缺失对致病性酵母光滑念珠菌细胞膜组成和物理化学性质的影响。具体而言,我们研究了选定固醇含量、磷脂组成、跨膜电位和PDR16基因活性的变化。使用高效液相色谱法测量固醇水平,通过薄层色谱法分析磷脂谱,用荧光光谱法评估跨膜电位,并用定量PCR测定基因表达水平。我们的研究结果显示,与野生型相比,Δerg6菌株中麦角固醇耗竭,酵母甾醇和羊毛甾醇含量增加,磷脂酰胆碱含量增加而磷脂酰乙醇胺含量减少。此外,Δerg6菌株表现出膜超极化,而PDR16表达没有变化。此外,Δerg6菌株对抗真菌药物米里霉素和金担子素A的敏感性增加。这些结果表明,ERG6基因缺失导致细胞膜组成发生显著改变,并可能在光滑念珠菌Δerg6缺失突变体中激活替代的麦角固醇合成途径。