Departamento de Microbiología y Genética/IBFG, Universidad de Salamanca/CSIC, Salamanca, Spain.
PLoS One. 2013 Aug 19;8(8):e71510. doi: 10.1371/journal.pone.0071510. eCollection 2013.
The regulation of cell wall synthesis by the clathrin light chain has been addressed. Schizosaccharomyces pombe clc1Δ mutant was inviable in the absence of osmotic stabilization; when grown in sorbitol-supplemented medium clc1Δ cells grew slowly, formed aggregates, and had strong defects in morphology. Additionally, clc1Δ cells exhibited an altered cell wall composition. A mutant that allowed modulating the amount of Clc1p was created to analyze in more detail the dependence of cell wall synthesis on clathrin. A 40% reduction in the amount of Clc1p did not affect acid phosphatase secretion and bulk lipid internalization. Under these conditions, β(1,3)glucan synthase activity and cell wall synthesis were reduced. Also, the delivery of glucan synthases to the cell surface, and the secretion of the Eng1p glucanase were defective. These results suggest that the defects in the cell wall observed in the conditional mutant were due to a defective secretion of enzymes involved in the synthesis/remodelling of this structure, rather than to their endocytosis. Our results show that a reduction in the amount of clathrin that has minor effects on general vesicle trafficking has a strong impact on cell wall synthesis, and suggest that this is the reason for the lethality of clc1Δ cells in the absence of osmotic stabilization.
网格蛋白轻链对细胞壁合成的调控作用已经得到解决。裂殖酵母 clc1Δ 突变体在没有渗透压稳定的情况下无法存活;当在添加山梨醇的培养基中生长时,clc1Δ 细胞生长缓慢,形成聚集体,并且在形态上有严重缺陷。此外,clc1Δ 细胞表现出改变的细胞壁组成。创建了一个可以调节 Clc1p 数量的突变体,以更详细地分析细胞壁合成对网格蛋白的依赖性。Clc1p 数量减少 40%不会影响酸性磷酸酶的分泌和大量脂质的内化。在这些条件下,β(1,3)葡聚糖合酶活性和细胞壁合成减少。此外,葡聚糖合酶向细胞表面的传递以及 Eng1p 葡聚糖酶的分泌也有缺陷。这些结果表明,条件性突变体中观察到的细胞壁缺陷是由于参与该结构合成/重塑的酶的分泌缺陷,而不是由于它们的内吞作用。我们的结果表明,网格蛋白数量的减少对一般囊泡运输的影响较小,但对细胞壁合成有很大的影响,并表明这是 clc1Δ 细胞在没有渗透压稳定的情况下致死的原因。