Lozančić Mateja, Žunar Bojan, Hrestak Dora, Lopandić Ksenija, Teparić Renata, Mrša Vladimir
Laboratory of Biochemistry, Faculty of Food Technology and Biotechnology, University of Zagreb, 10000 Zagreb, Croatia.
Department of Biotechnology, University of Natural Resources and Applied Life Sciences, 1180 Vienna, Austria.
J Fungi (Basel). 2021 Feb 9;7(2):128. doi: 10.3390/jof7020128.
Yeast cell walls have two major roles, to preserve physical integrity of the cell, and to ensure communication with surrounding molecules and cells. While the first function requires evolutionary conserved polysaccharide network synthesis, the second needs to be flexible and provide adaptability to different habitats and lifestyles. In this study, the comparative analysis of proteins required for cell wall biosynthesis and functions containing 187 proteins of 92 different yeasts was performed in order to assess which proteins were broadly conserved among yeasts and which were more species specific. Proteins were divided into several groups according to their role and localization. As expected, many proteins involved in protein glycosylation, glycosylphosphatidylinositol (GPI) synthesis and the synthesis of wall polysaccharides had orthologues in most other yeasts. Similarly, a group of GPI anchored proteins involved in cell wall biosynthesis (Gas proteins and Dfg5p/Dcw1p) and other non-GPI anchored cell wall proteins involved in the wall synthesis and remodeling were highly conserved. However, GPI anchored proteins involved in flocculation, aggregation, cell separation, and those of still unknown functions were not highly conserved. The proteins localized in the cell walls of various yeast species were also analyzed by protein biotinylation and blotting. Pronounced differences were found both in the patterns, as well as in the overall amounts of different groups of proteins. The amount of GPI-anchored proteins correlated with the mannan to glucan ratio of the wall. Changes of the wall proteome upon temperature shift to 42 °C were detected.
酵母细胞壁有两个主要作用,一是维持细胞的物理完整性,二是确保与周围分子和细胞进行通讯。虽然第一个功能需要进化上保守的多糖网络合成,但第二个功能需要具有灵活性,并能适应不同的栖息地和生活方式。在本研究中,对92种不同酵母的187种参与细胞壁生物合成和功能的蛋白质进行了比较分析,以评估哪些蛋白质在酵母中广泛保守,哪些更具物种特异性。根据蛋白质的作用和定位将其分为几组。正如预期的那样,许多参与蛋白质糖基化、糖基磷脂酰肌醇(GPI)合成和细胞壁多糖合成的蛋白质在大多数其他酵母中都有直系同源物。同样,一组参与细胞壁生物合成的GPI锚定蛋白(Gas蛋白和Dfg5p/Dcw1p)以及其他参与细胞壁合成和重塑的非GPI锚定细胞壁蛋白高度保守。然而,参与絮凝、聚集、细胞分离的GPI锚定蛋白以及功能仍未知的GPI锚定蛋白并不高度保守。还通过蛋白质生物素化和印迹分析了定位于各种酵母细胞壁中的蛋白质。在不同蛋白质组的模式以及总量上都发现了明显差异。GPI锚定蛋白的量与细胞壁中甘露聚糖与葡聚糖的比例相关。检测到温度转移至42°C时细胞壁蛋白质组的变化。