糖基磷脂酰肌醇(GPI)附着蛋白编码基因的上调对酿酒酵母FKS1破坏引起的细胞壁损伤的响应

H Terashima, N Yabuki, M Arisawa, K Hamada, K Kitada
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引用次数: 86

摘要

FKS1和FKS2编码葡聚糖合成酶的替代催化亚基,负责在酿酒酵母细胞壁中合成β -1,3-葡聚糖。FKS1的破坏降低了细胞壁的葡聚糖含量,增加了几丁质含量,并激活了CWP1的表达,CWP1编码糖基磷脂酰肌醇(GPI)依赖性细胞壁蛋白。这些细胞反应被认为是对细胞壁损伤的补偿,以保持细胞壁的完整性。在这里,我们报告了鉴定,通过全基因组筛选,22个基因在fks1delta细胞中转录上调。其中,发现5个基因编码gpi附着蛋白,其中3个与细胞壁共价相关。对启动子区域的缺失和替换分析发现,rlm1结合序列是FKS1断裂后上调的原因。利用强力霉素可以抑制FKS1表达的rlm1delta tetOp-FKS1菌株,我们检测了Rlm1对这五个基因转录上调的要求。这5个基因中有3个未被强力霉素上调,表明当FKS1失活时,Rlm1介导了它们的上调。其余两个基因被强力霉素上调,这表明除了Rlm1外,还有一个转录因子参与了它们对FKS1破坏的反应。
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Up-regulation of genes encoding glycosylphosphatidylinositol (GPI)-attached proteins in response to cell wall damage caused by disruption of FKS1 in Saccharomyces cerevisiae.

FKS1 and FKS2 encode alternative catalytic subunits of the glucan synthases that are responsible for synthesis of beta-1,3-glucan in the Saccharomyces cerevisiae cell wall. Disruption of FKS1 reduces the glucan content of the cell wall, increases chitin content and activates the expression of CWP1, which encodes a glycosylphosphatidylinositol (GPI)-dependent cell wall protein. These cellular responses have been regarded as compensating for cell wall damage in order to maintain cell wall integrity. Here, we report the identification, by genome-wide screening, of 22 genes that are transcriptionally up-regulated in fks1delta cells. Among them, five genes were found to encode GPI-attached proteins, three of which are covalently associated with the cell wall. Deletion and replacement analysis of the promoter regions identified Rlm1-binding sequences as being responsible for the up-regulation following disruption of FKS1. Using the rlm1delta tetOp-FKS1 strain, in which the expression of FKS1 can be repressed by doxycycline, we examined the requirement for Rlm1 for the transcriptional up-regulation of these five genes. Three of the five genes were not up-regulated by doxycycline, indicating that Rlm1 mediates their up-regulation when FKS1 is inactivated. The remaining two genes were up-regulated by doxycycline, suggesting that a transcription factor other than Rlm1 is involved in their response to disruption of FKS1.

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