Benzyl Alcohol Increases Diffusion Limit of Nuclear Membrane in Saccharomyces cerevisiae Cells

Abstract

Objective: Fungi are invasive species responsible for infections in many people around the world and which severely affect the immune system. The opportunistic pathogenic species, such as Candida species and Aspergillus fumigatus, can cause death in people with weakened immune systems. Natural medicines derived from plants are often used to treat fungal diseases. In connection with our efforts to unearth possible cellular targets of antimicrobial agents, in this study, we aimed to determine the functional consequences of benzyl alcohol treatment on the nuclear membrane. Materials and Methods: We analysed the nuclear membrane distortions caused by benzyl alcohol in Saccharomyces cerevisiae cells using Nup49-GFP reporter strain. We also studied cellular distributions of various fluorescently tagged nuclearcytoplasmic shuttling proteins to determine any functional disturbances in nuclear pore complexes upon benzyl alcohol treatment. Localization of 51.5 kDa protein LexA-NES-GFP and 61.8 kDa protein Pho4(Δ157-164)-GFP to the nucleus in yeast cells was key for evaluating the effect upon diffusion limit of pores. Results: By analyzing the distribution of fluorescently tagged nuclear localization signal or nuclear export signals bearing reporter proteins between the nucleus and cytoplasm, we have shown that the nuclear membrane becomes leaky upon benzyl alcohol treatment. Conclusion: The diffusion limit across the nuclear membrane in yeast cells is increased upon benzyl alcohol treatment. We believe that these findings not only increase our understanding of the mode of action of benzyl alcohol bearing antifungal agents, but also help widening their use.