title: Yeast CUP1 protects HeLa cells against copper-induced stress.
abstract: As an essential trace element, copper can be toxic in mammalian cells when present in excess. Metallothioneins (MTs) are small, cysteine-rich proteins that avidly bind copper and thus play an important role in detoxification. Yeast CUP1 is a member of the MT gene family. The aim of this study was to determine whether yeast CUP1 could bind copper effectively and protect cells against copper stress. In this study, CUP1 expression was determined by quantitative real-time PCR, and copper content was detected by inductively coupled plasma mass spectrometry. Production of intracellular reactive oxygen species (ROS) was evaluated using the 2',7'-dichlorofluorescein-diacetate (DCFH-DA) assay. Cellular viability was detected using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, and the cell cycle distribution of CUP1 was analyzed by fluorescence-activated cell sorting. The data indicated that overexpression of yeast CUP1 in HeLa cells played a protective role against copper-induced stress, leading to increased cellular viability (P<0.05) and decreased ROS production (P<0.05). It was also observed that overexpression of yeast CUP1 reduced the percentage of G1 cells and increased the percentage of S cells, which suggested that it contributed to cell viability. We found that overexpression of yeast CUP1 protected HeLa cells against copper stress. These results offer useful data to elucidate the mechanism of the MT gene on copper metabolism in mammalian cells.
authors.affiliation: School of Agriculture and Biology, Department of Animal Sciences, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: X X
authors.initials: XX
authors.lastname: Xie
authors.affiliation: School of Agriculture and Biology, Department of Animal Sciences, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Y F
authors.initials: YF
authors.lastname: Ma
authors.affiliation: School of Agriculture and Biology, Department of Animal Sciences, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Q S
authors.initials: QS
authors.lastname: Wang
authors.affiliation: School of Agriculture and Biology, Department of Animal Sciences, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Z L
authors.initials: ZL
authors.lastname: Chen
authors.affiliation: School of Agriculture and Biology, Department of Animal Sciences, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: R R
authors.initials: RR
authors.lastname: Liao
authors.affiliation: School of Agriculture and Biology, Department of Animal Sciences, Shanghai Jiao Tong University, Shanghai, China.
authors.forename: Y C
authors.initials: YC
authors.lastname: Pan
mesh_terms: D002352:Carrier Proteins / Q000032:analysis / Q000378:metabolism; D002453:Cell Cycle / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D003300:Copper / Q000032:analysis / Q000378:metabolism*; D005562:Formazans; D006367:HeLa Cells; D006801:Humans; D008668:Metallothionein / Q000032:analysis / Q000502:physiology*; D018384:Oxidative Stress / Q000502:physiology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D013778:Tetrazolium Salts; D013997:Time Factors
chemical_list: C500769:CUP1-1 protein, S cerevisiae; D002352:Carrier Proteins; D005562:Formazans; D017382:Reactive Oxygen Species; D013778:Tetrazolium Salts; C079782:MTT formazan; D003300:Copper; D008668:Metallothionein