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必需蛋白fap7参与酿酒酵母的氧化应激反应。

The essential protein fap7 is involved in the oxidative stress response of Saccharomyces cerevisiae.

作者信息

Juhnke H, Charizanis C, Latifi F, Krems B, Entian K D

机构信息

Institut für Mikrobiologie der Johann Wolfgang Goethe-Universität Frankfurt, Biozentrum, Niederursel, Marie-Curie-Strasse 9, D-60439 Frankfurt am Main, Germany.

出版信息

Mol Microbiol. 2000 Feb;35(4):936-48. doi: 10.1046/j.1365-2958.2000.01768.x.

Abstract

Pos9 (Skn7) is an important transcription factor that, together with Yap1, induces the expression of oxidative stress target genes in Saccharomyces cerevisiae. The activation of Pos9 upon an oxidative stress signal occurs post-translationally. In a mutant screen for factors involved in the activation of a Pos9-dependent reporter gene upon oxidative stress, we identified the mutant fap7-1 (for factor activating Pos9). This point mutant failed to activate a Gal4-Pos9 hybrid transcription factor, assayed by hydrogen peroxide-induced GAL1-lacZ reporter gene activities. Additionally, the fap7-1 mutant strain was sensitive to oxidative stress and revealed slow growth on glucose compared with the wild type. The fap7-1 mutation also affected the induction of the Pos9 target gene TPX1 and of a synthetic promoter previously identified to be regulated in a Yap1- and Pos9-dependent manner. This lack of induction was specific as the fap7-1 mutant response to other stresses such as sodium chloride or co-application of both hydrogen peroxide and sodium chloride was not affected, as tested with the Pos9-independent expression pattern of a TPS2-lacZ reporter system. We identified the gene YDL166c to be allelic to the FAP7 gene and to be essential. Fluorescence microscopy of Fap7-GFP fusion proteins indicated a nuclear localization of the Fap7 protein. Our data suggest that Fap7 is a nuclear factor important for Pos9-dependent target gene transcription upon oxidative stress.

摘要

Pos9(Skn7)是一种重要的转录因子,它与Yap1一起在酿酒酵母中诱导氧化应激靶基因的表达。氧化应激信号作用下Pos9的激活发生在翻译后。在一个针对氧化应激时参与激活Pos9依赖性报告基因的因子的突变体筛选中,我们鉴定出了突变体fap7-1(因子激活Pos9)。通过过氧化氢诱导的GAL1-lacZ报告基因活性检测,这个点突变体无法激活Gal4-Pos9杂交转录因子。此外,fap7-1突变体菌株对氧化应激敏感,与野生型相比,在葡萄糖上生长缓慢。fap7-1突变还影响了Pos9靶基因TPX1以及先前鉴定为以Yap1和Pos9依赖性方式调控的合成启动子的诱导。这种诱导缺失是特异性的,因为用TPS2-lacZ报告系统的Pos9非依赖性表达模式测试发现,fap7-1突变体对其他应激(如氯化钠或过氧化氢和氯化钠共同作用)的反应不受影响。我们鉴定出基因YDL166c与FAP7基因等位且是必需的。Fap7-GFP融合蛋白的荧光显微镜观察表明Fap7蛋白定位于细胞核。我们的数据表明,Fap7是一种在氧化应激时对Pos9依赖性靶基因转录很重要的核因子。

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