UV and arsenate toxicity: a specific and sensitive yeast bioluminescence assay

被引:18
作者
Bakhrat, Anya [1 ]
Eltzov, Evgeni [2 ]
Finkelstein, Yishay [1 ]
Marks, Robert S. [3 ,4 ,5 ]
Raveh, Dina [1 ]
机构
[1] Ben Gurion Univ Negev, Dept Life Sci, Fac Nat Sci, IL-84105 Beer Sheva, Israel
[2] Ben Gurion Univ Negev, Unit Environm Engn, IL-84105 Beer Sheva, Israel
[3] Ben Gurion Univ Negev, Dept Biotechnol Engn, Fac Engn Sci, IL-84105 Beer Sheva, Israel
[4] Ben Gurion Univ Negev, Natl Inst Biotechnol Negev, IL-84105 Beer Sheva, Israel
[5] Ben Gurion Univ Negev, Ctr Meso & Nanoscale Sci & Technol, IL-84105 Beer Sheva, Israel
关键词
Bacterial luciferase; F-box protein; UV; Sunscreen; Arsenate; DNA-DAMAGE RESPONSE; DEAD-SEA MINERALS; UBA PROTEIN DDI1; SACCHAROMYCES-CEREVISIAE; HO ENDONUCLEASE; BACTERIAL BIOLUMINESCENCE; ENVIRONMENTAL-CHANGES; EXPRESSION; TRANSCRIPTION; DEGRADATION;
D O I
10.1007/s10565-011-9184-8
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
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.
引用
收藏
页码:227 / 236
页数:10
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