Monitoring Oxidative Stress and DNA Damage Induced by Heavy Metals in Yeast Expressing a Redox-Sensitive Green Fluorescent Protein |
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Authors: | Shanshan Yu Wei Qin Guoqiang Zhuang Xianen Zhang Guanjun Chen Weifeng Liu |
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Institution: | (1) The State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, No. 27 Shanda South Road, Jinan, 250100, Shandong, People’s Republic of China;(2) Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, 18 Shuangqing Road, Haidian District, Beijing, 10085, People’s Republic of China;(3) Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan, 430071, Hubei, People’s Republic of China |
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Abstract: | Reactive oxygen species (ROS) has been proposed to play an important role in heavy metal-associated toxicity and pathology.
Conventional methods for determining ambient redox state in cells are usually labor-intensive, precluding real-time or single-cell
monitoring changes in intracellular redox poise resulting from either metabolic processes or environmental influences. Redox-sensitive
green fluorescent protein (roGFP) was expressed in Saccharomyces cerevisiae and recombinant cells were evaluated in monitoring the changes in the redox state of living cells when challenged with toxicologically
relevant metal ions. roGFP expressed in yeast responded not only to typical membrane-permeant oxidants and reductants, but
also to toxicological metal ion-induced intracellular redox changes. Moreover, exposure of yeast cells to NaAsO2 or Pb(NO3)2 at concentrations that induced redox changes reported by roGFP caused up to two- to three-fold increases in DNA mutation
frequency. This mutagenic effect was largely caused by oxidative stress since blocking the production of hydryl radicals significantly
reduced the mutation rate as well as delayed the cell death. |
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