UV and arsenate toxicity: a specific and sensitive yeast bioluminescence assay
UV and arsenate toxicity: a specific and sensitive yeast bioluminescence assay
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DOI：
10.1007/s10565-011-9184-8
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发表时间：
2011-06-01
影响因子：
6.1
通讯作者：
Raveh, Dina
中科院分区：
文献类型：
--
作者：
Bakhrat, Anya;Eltzov, Evgeni;Raveh, Dina
我们描述了一种酿酒酵母对紫外线和砷盐的生物发光分析方法，其中细菌荧光素酶基因受酵母基因UFO1启动子的调控。UFO1编码Skp1-CDC53-F-box蛋白泛素连接酶复合体的F-box亚基，受DNA损伤和砷盐诱导。我们将UFO1启动子改造成一个现有的酵母生物报告程序，它使用人类基因来检测水体中的类固醇激素干扰化合物。我们的分析表明，在不同的突变背景下使用内源酵母启动子可以区分不同的环境信号。UFO1工程酵母对UVB有很强的生物发光响应，可用于评价紫外线防护防晒霜。它们在检测极低浓度的砷酸盐方面也很有效，特别是在缺乏释放有毒化学物质机制的pdr5 Delta突变体中；然而，它们对镉或汞没有反应。结合使用内源酵母启动子元件和胁迫反应途径的突变体可以促进高特异性酵母生物记者的开发，从而能够区分环境中存在的密切相关的化学物质。
We describe a Saccharomyces cerevisiae bioluminescence assay for UV and arsenate in which bacterial luciferase genes are regulated by the promoter of the yeast gene, UFO1. UFO1 encodes the F-box subunit of the Skp1-Cdc53-F-box protein ubiquitin ligase complex and is induced by DNA damage and by arsenate. We engineered the UFO1 promoter into an existing yeast bioreporter that employs human genes for detection of steroid hormone-disrupting compounds in water bodies. Our analysis indicates that use of an endogenous yeast promoter in different mutant backgrounds allows discrimination between different environmental signals. The UFO1-engineered yeast give a robust bioluminescence response to UVB and can be used for evaluating UV protective sunscreens. They are also effective in detecting extremely low concentrations of arsenate, particularly in pdr5 Delta mutants that lack a mechanism to extrude toxic chemicals; however, they do not respond to cadmium or mercury. Combined use of endogenous yeast promoter elements and mutants of stress response pathways may facilitate development of high-specificity yeast bioreporters able to discriminate between closely related chemicals present together in the environment.