Université de Toulouse, INSA, UPS, INP, 135 avenue de Rangueil, 31077, Toulouse, France,
Curr Genet. 2013 Nov;59(4):187-96. doi: 10.1007/s00294-013-0411-0. Epub 2013 Sep 27.
Over the past 20 years, the yeast cell wall has been thoroughly investigated by genetic and biochemical methods, leading to remarkable advances in the understanding of its biogenesis and molecular architecture as well as to the mechanisms by which this organelle is remodeled in response to environmental stresses. Being a dynamic structure that constitutes the frontier between the cell interior and its immediate surroundings, imaging cell surface, measuring mechanical properties of cell wall or probing cell surface proteins for localization or interaction with external biomolecules are among the most burning questions that biologists wished to address in order to better understand the structure-function relationships of yeast cell wall in adhesion, flocculation, aggregation, biofilm formation, interaction with antifungal drugs or toxins, as well as response to environmental stresses, such as temperature changes, osmotic pressure, shearing stress, etc. The atomic force microscopy (AFM) is nowadays the most qualified and developed technique that offers the possibilities to address these questions since it allows working directly on living cells to explore and manipulate cell surface properties at nanometer resolution and to analyze cell wall proteins at the single molecule level. In this minireview, we will summarize the most recent contributions made by AFM in the analysis of the biomechanical and biochemical properties of the yeast cell wall and illustrate the power of this tool to unravel unexpected effects caused by environmental stresses and antifungal agents on the surface of living yeast cells.
在过去的 20 年中,通过遗传和生化方法对酵母细胞壁进行了深入研究,这使得人们对其生物发生和分子结构以及细胞器响应环境压力进行重塑的机制有了显著的认识。作为构成细胞内部与其周围环境之间边界的动态结构,对细胞表面进行成像、测量细胞壁的机械性能、探测细胞表面蛋白的定位或与外部生物分子的相互作用,是生物学家为了更好地理解酵母细胞壁在黏附、絮凝、聚集、生物膜形成、与抗真菌药物或毒素相互作用以及对温度变化、渗透压、剪切力等环境压力的反应中的结构-功能关系而希望解决的最热门问题之一。原子力显微镜(AFM)是目前最有资格和最先进的技术,它提供了研究这些问题的可能性,因为它可以直接在活细胞上工作,以纳米分辨率探索和操纵细胞表面特性,并在单分子水平上分析细胞壁蛋白。在这篇综述中,我们将总结 AFM 在分析酵母细胞壁的生物力学和生化特性方面的最新贡献,并说明该工具在揭示环境压力和抗真菌剂对活酵母细胞表面的意外影响方面的强大功能。