The role of MT (metallothionein) gene expression was investigated in rotenone-treated HeLa cells to induce a deficiency of NADH:ubiquinone oxidoreductase (complex I). Complex I deficiency leads to a diversity of cellular consequences, including production of ROS (reactive oxygen species) and apoptosis. HeLa cells were titrated with rotenone, resulting in dose-dependent decrease in complex I activity and elevated ROS production at activities lower than 33%. Expression of MT2A (MT isoform 2A), but not MT1A or MT1B RNA, was significantly inducible by rotenone (up to 7-fold), t-BHP (t-butyl hydroperoxide; 5-fold) and CdCl2 (50-fold), but not ZnCl2. Myxothiazol treatment did not elevate either ROS or MT2A levels, which supports a ROS-related mechanism for rotenone-induced MT2A expression. To evaluate the role of MT2A expression, MT2A and MT1B were overexpressed in HeLa cells and treated with rotenone. Compared with control and MT1B-overexpressing cells, ROS production was significantly lower and cell viability higher in MT2A-overexpressing HeLa cells when ROS production was enhanced by treatment with t-BHP. Mitochondrial membrane potential was noticeably less reduced in both MT-overexpressing cell lines. MT2A overexpression in rotenone-treated cells also significantly reduced or delayed apoptosis induction, as measured by caspase 3/7 activity and cytosolic nucleosome enrichment. We conclude that MT2A offers significant protection against the main death-causing consequences of rotenone-induced complex I deficiency in HeLa cells. Our results are in support of the protective role against oxidative stress ascribed to MTs and provide evidence that MT2A expression may be a beneficial downstream adaptive response in complex I-deficient cells.
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Research Article|
March 28 2006
Metallothionein isoform 2A expression is inducible and protects against ROS-mediated cell death in rotenone-treated HeLa cells
Fimmie Reinecke;
Fimmie Reinecke
*Division of Biochemistry, School for Chemistry and Biochemistry, North-West University, Potchefstroom 2531, South Africa
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Oksana Levanets;
Oksana Levanets
*Division of Biochemistry, School for Chemistry and Biochemistry, North-West University, Potchefstroom 2531, South Africa
†Institute of Molecular Biology and Genetics of the National Academy of Sciences of Ukraine, Kiev 03143, Ukraine
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Yolanda Olivier;
Yolanda Olivier
*Division of Biochemistry, School for Chemistry and Biochemistry, North-West University, Potchefstroom 2531, South Africa
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Roan Louw;
Roan Louw
*Division of Biochemistry, School for Chemistry and Biochemistry, North-West University, Potchefstroom 2531, South Africa
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Boitumelo Semete;
Boitumelo Semete
‡Centre for Genome Research, North-West University, Pretoria 0020, South Africa
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Anne Grobler;
Anne Grobler
§School for Pharmacy, North-West University, Potchefstroom 2531, South Africa
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Juan Hidalgo;
Juan Hidalgo
∥Institute of Neurosciences, Department of Cellular Biology, Physiology and Immunology, Faculty of Sciences, Autonomous University of Barcelona, Barcelona 08193, Spain
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Jan Smeitink;
Jan Smeitink
¶Nijmegen Centre for Mitochondrial Disorders, Radboud University Nijmegen Medical Centre, Nijmegen 6500, The Netherlands
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Antonel Olckers;
Antonel Olckers
‡Centre for Genome Research, North-West University, Pretoria 0020, South Africa
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Francois H. Van Der Westhuizen
Francois H. Van Der Westhuizen
1
*Division of Biochemistry, School for Chemistry and Biochemistry, North-West University, Potchefstroom 2531, South Africa
1To whom correspondence should be addressed (email bchfhvdw@puk.ac.za).
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Publisher: Portland Press Ltd
Received:
August 02 2005
Revision Received:
December 20 2005
Accepted:
January 10 2006
Accepted Manuscript online:
January 10 2006
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London
2006
Biochem J (2006) 395 (2): 405–415.
Article history
Received:
August 02 2005
Revision Received:
December 20 2005
Accepted:
January 10 2006
Accepted Manuscript online:
January 10 2006
Citation
Fimmie Reinecke, Oksana Levanets, Yolanda Olivier, Roan Louw, Boitumelo Semete, Anne Grobler, Juan Hidalgo, Jan Smeitink, Antonel Olckers, Francois H. Van Der Westhuizen; Metallothionein isoform 2A expression is inducible and protects against ROS-mediated cell death in rotenone-treated HeLa cells. Biochem J 15 April 2006; 395 (2): 405–415. doi: https://doi.org/10.1042/BJ20051253
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