Laboratory of Microbial Genetics, Department of Genetics, Federal University of Pernambuco, Recife, Brazil.
Laboratory of Glycobiology, Centre for Organismal Studies, University of Heidelberg, Heidelberg, Germany.
Antonie Van Leeuwenhoek. 2021 Jul;114(7):1141-1153. doi: 10.1007/s10482-021-01584-w. Epub 2021 May 4.
The NCW2 gene was recently described as encoding a GPI-bounded protein that assists in the re-modelling of the Saccharomyces cerevisiae cell wall (CW) and in the repair of damage caused by the polyhexamethylene biguanide (PHMB) polymer to the cell wall. Its absence produces a re-organization of the CW structure that result in resistance to lysis by glucanase. Hence, the present study aimed to extend the analysis of the Ncw2 protein (Ncw2p) to determine its physiological role in the yeast cell surface. The results showed that Ncw2p is transported to the cell surface upon O-mannosylation mediated by the Pmt1p-Pmt2p enzyme complex. It co-localises with the yeast bud scars, a region in cell surface formed by chitin deposition. Once there, Ncw2p enables correct chitin/β-glucan structuring during the exponential growth. The increase in molecular mass by hyper-mannosylation coincides with the increasing in chitin deposition, and leads to glucanase resistance. Treatment of the yeast cells with PHMB produced the same biological effects observed for the passage from exponential to stationary growth phase. This might be a possible mechanism of yeast protection against cationic biocides. In conclusion, we propose that Ncw2p takes part in the mechanism involved in the control of cell surface rigidity by aiding on the linkage between chitin and glucan layers in the modelling of the cell wall during cell growth.
NCW2 基因最近被描述为编码一种 GPI 结合蛋白,该蛋白有助于重塑酿酒酵母细胞壁 (CW),并修复多聚六亚甲基双胍 (PHMB) 聚合物对细胞壁造成的损伤。该基因的缺失会导致 CW 结构重新组织,从而使细胞对葡聚糖酶的裂解产生抗性。因此,本研究旨在扩展对 Ncw2 蛋白 (Ncw2p) 的分析,以确定其在酵母细胞表面的生理作用。结果表明,Ncw2p 是通过 Pmt1p-Pmt2p 酶复合物介导的 O-甘露糖化而被转运到细胞表面的。它与酵母芽痕共定位,芽痕是由几丁质沉积形成的细胞表面区域。一旦到达那里,Ncw2p 就能够在指数生长期正确构建几丁质/β-葡聚糖结构。通过过度甘露糖化增加分子量与几丁质沉积的增加相吻合,并导致对葡聚糖酶的抗性。用 PHMB 处理酵母细胞会产生与从指数生长期到静止生长期的转变所观察到的相同的生物学效应。这可能是酵母对抗阳离子杀生物剂的保护的一种可能机制。总之,我们提出 Ncw2p 参与了控制细胞表面刚性的机制,通过协助细胞生长过程中细胞壁建模中几丁质和葡聚糖层之间的连接,从而参与其中。