KEG1/YFR042w encodes a novel Kre6-binding endoplasmic reticulum membrane protein responsible for β-1,6-glucan synthesis in Saccharomyces cerevisiae
KEG1/YFR042w encodes a novel Kre6-binding endoplasmic reticulum membrane protein responsible for β-1,6-glucan synthesis in Saccharomyces cerevisiae
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DOI：
10.1074/jbc.m706486200
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发表时间：
2007-11-23
影响因子：
4.8
通讯作者：
Yoda, Koji
中科院分区：
文献类型：
--
作者：
Nakamata, Kosuke;Kurita, Tomokazu;Yoda, Koji
酿酒酵母的 KEG1/YFR042w 是一个必需基因，编码 200 个氨基酸的多肽，具有四个预测的跨膜结构域。绿色荧光蛋白或Myc(6)标记的Keg1蛋白显示出整合膜蛋白的典型特征，并通过荧光成像在内质网中发现。 Triton X-100 溶解的细胞裂解液的免疫沉淀显示，Keg1 与 Kre6 结合，已知 Kre6 参与 β-1,6-葡聚糖的合成。为了更详细地分析 Keg1 的基本功能，我们通过易错聚合酶链式反应构建了温度敏感突变等位基因。 keg1-1突变细胞表现出与Delta kre6突变体相同的表型，包括对Calcoflor白过敏、对K1杀伤毒素敏感性降低以及细胞壁中β-1,6-葡聚糖含量降低。这些结果表明，Keg1 和 Kre6 在酿酒酵母的 β-1,6-葡聚糖合成中具有协同作用。
KEG1/YFR042w of Saccharomyces cerevisiae is an essential gene that encodes a 200-amino acid polypeptide with four predicted transmembrane domains. The green fluorescent protein or Myc(6)-tagged Keg1 protein showed the typical characteristics of an integral membrane protein and was found in the endoplasmic reticulum by fluorescence imaging. Immunoprecipitation from the Triton X-100-solubilized cell lysate revealed that Keg1 binds to Kre6, which has been known to participate in beta-1,6-glucan synthesis. To analyze the essential function of Keg1 in more detail, we constructed temperature-sensitive mutant alleles by error-prone polymerase chain reaction. The keg1-1 mutant cells showed a common phenotype with Delta kre6 mutant including hypersensitivity to Calcofluor white, reduced sensitivity to the K1 killer toxin, and reduced content of beta-1,6-glucan in the cell wall. These results suggest that Keg1 and Kre6 have a cooperative role in beta-1,6-glucan synthesis in S. cerevisiae.