Department of Life Science, College of Science, Rikkyo University, Tokyo, Japan.
Graduate School of Science, Osaka Metropolitan University, Osaka, Japan.
Microbiologyopen. 2023 Oct;12(5):e1385. doi: 10.1002/mbo3.1385.
Peptidoglycan for elongation in Escherichia coli is synthesized by the Rod complex, which includes RodZ. Although various mutant strains of the Rod complex have been isolated, the relationship between the activity of the Rod complex and the overall physical and chemical structures of the peptidoglycan have not been reported. We constructed a RodZ mutant, termed RMR, and analyzed the growth rate, morphology, and other characteristics of cells producing the Rod complexes containing RMR. The growth and morphology of RMR cells were abnormal, and we isolated suppressor mutants from RMR cells. Most of the suppressor mutations were found in components of the Rod complex, suggesting that these suppressor mutations increase the integrity and/or the activity of the Rod complex. We purified peptidoglycan from wild-type, RMR, and suppressor mutant cells and observed their structures in detail. We found that the peptidoglycan purified from RMR cells had many large holes and different compositions of muropeptides from those of WT cells. The Rod complex may be a determinant not only for the whole shape of peptidoglycan but also for its highly dense structure to support the mechanical strength of the cell wall.
在大肠杆菌中,肽聚糖的延伸是由 Rod 复合物合成的,其中包括 RodZ。尽管已经分离出各种 Rod 复合物的突变株,但 Rod 复合物的活性与肽聚糖的整体物理化学结构之间的关系尚未报道。我们构建了一个 RodZ 突变体,称为 RMR,并分析了含有 RMR 的 Rod 复合物产生的细胞的生长速率、形态和其他特征。RMR 细胞的生长和形态异常,我们从 RMR 细胞中分离出了抑制子突变体。大多数抑制子突变发生在 Rod 复合物的成分中,这表明这些抑制子突变增加了 Rod 复合物的完整性和/或活性。我们从野生型、RMR 和抑制子突变体细胞中纯化了肽聚糖,并详细观察了它们的结构。我们发现,从 RMR 细胞中纯化的肽聚糖有许多大孔,其寡肽组成与 WT 细胞不同。Rod 复合物可能不仅是决定肽聚糖整体形状的因素,也是决定其高密度结构以支撑细胞壁机械强度的因素。