A single FKS homologue in Yarrowia lipolytica is essential for viability
A single FKS homologue in Yarrowia lipolytica is essential for viability
复制标题
解脂耶氏酵母中的单个 FKS 同源物对于活力至关重要
作者：
M. León;R. Sentandreu;J. Zueco
β-1,3-葡聚糖是酵母细胞壁的结构成分，赋予细胞形状，其合成发生在质膜上，是至少一个双成分复合物活性的结果。 Fks1p 是直接负责 β-1,3-葡聚糖合成的催化亚基，而第二个亚基 Rho1p 具有 GTP 依赖性调节作用。 FKS1 已在酿酒酵母中得到表征，其功能与 FKS2/GSC2 的功能至少部分冗余。 FKS 同源物也在其他几种真菌物种中被发现，包括白色念珠菌、粟酒裂殖酵母、构巢曲霉、新型隐球菌和巴西副球孢子菌。在这项工作中，我们使用源自 Fks1ps 保守区域的简并寡核苷酸来分离严格需氧非传统酵母解脂耶氏酵母可能的 FKS 同源基因。使用这种方法，我们分离出了一个 FKS 同源物，我们将其命名为 YlFKS1；它编码一个 1961 个氨基酸的蛋白质，与其他 Fksps 表现出高度的同源性。 YlFKS1在影响细胞壁的不同条件下的表达分析没有显示出显着差异。最后，尽管使用了两种不同的技术，但获得含有破坏的 YlFKS1 等位基因的解脂耶氏酵母菌株的尝试还是失败了。综上所述，这些结果表明，与酿酒酵母不同，YlFKS1 是解脂耶氏酵母中唯一的 FKS1 同源物，并且对于生长至关重要。版权所有 © 2002 约翰威利父子有限公司
The synthesis of β‐1,3‐glucan, the structural component of the yeast cell wall which gives shape to the cell, occurs at the plasma membrane and is the result of the activity of at least a two‐component complex. Fks1p is the catalytic subunit directly responsible for the synthesis of β‐1,3‐glucan, whilst the second subunit, Rho1p, has a GTP‐dependent regulatory role. FKS1 has been characterized in Saccharomyces cerevisiae, where its function is at least partially redundant with that of FKS2/GSC2. FKS homologues have also been identified in several other fungal species, including Candida albicans, Schizosaccharomyces pombe, Aspergillus nidulans, Cryptococcus neoformans and Paracoccidiodes brasiliensis. In this work, we have used degenerate oligonucleotides derived from the conserved regions of Fks1ps to isolate the possible FKS homologue genes of the strictly aerobic non‐conventional yeast Yarrowia lipolytica. Using this approach, we have isolated a single FKS homologue which we have named YlFKS1; this codes a 1961 amino acid protein that shows a high degree of homology with other Fksps. Expression analysis of YlFKS1 under different conditions affecting the cell wall did not reveal significant differences. Finally, attempts to obtain a Y. lipolytica strain containing a disrupted YlFKS1 allele failed, despite having used two different techniques. Taken together, these results suggest that, unlike S. cerevisiae, YlFKS1 is the only FKS1 homologue in Y. lipolytica and is essential for growth. Copyright © 2002 John Wiley & Sons, Ltd.