Blagoeva J, Stoev G, Venkov P
Institute of Molecular Biology, Bulgarian Academy of Sciences, Sofia.
Yeast. 1991 Jul;7(5):455-61. doi: 10.1002/yea.320070504.
The phenotype of VY1160 fragile Saccharomyces cerevisiae mutant is characterized by cell lysis upon transfer to hypotonic solutions and increased permeability of cells growing in osmotically stabilized media. Two mutations, srb1 and ts1, have been identified in VY1160 cells and previous studies have shown that the increased permeability is due to the ts1 mutation which causes a shortening of mannan side-chains. Here we report that the srb1 mutation, which is the genetic determinant of cell lysis, is responsible for quantitative and structural changes of glucans. Experiments with isogenic single mutation strains, genetic studies coupled with quantitative measurements of glucan content per cell, and methylation analysis of glucans provide evidence that srb1 mutation leads to i) formation of mechanically unstable cell wall network made of insoluble glucan fibrils which are shorter and contain beta(1-6) inter-residue linkages and ii) insufficient filling of the space between the fibrils due to a shortage of the alkali-soluble glucan. Although growing exponentially in osmotically stabilized media, the srb1 cells cannot resist an osmotic shock and, hence, burst immediately.
VY1160脆弱型酿酒酵母突变体的表型特征为:转移至低渗溶液后细胞裂解,以及在渗透稳定培养基中生长的细胞通透性增加。在VY1160细胞中已鉴定出两个突变,即srb1和ts1,先前的研究表明,通透性增加是由于ts1突变导致甘露聚糖侧链缩短所致。在此我们报告,作为细胞裂解的遗传决定因素,srb1突变导致葡聚糖的数量和结构发生变化。利用同基因单突变菌株进行的实验、结合对每个细胞葡聚糖含量的定量测量的遗传学研究以及葡聚糖的甲基化分析均提供了证据,表明srb1突变导致:i) 由不溶性葡聚糖原纤维构成的机械不稳定细胞壁网络的形成,这些原纤维较短且含有β(1-6)残基间连接;ii) 由于碱溶性葡聚糖短缺,原纤维之间的空间填充不足。尽管srb1细胞在渗透稳定培养基中呈指数生长,但它们无法抵抗渗透压冲击,因此会立即破裂。