Nuclear and mitochondrial genome responses in HeLa cells treated with inhibitors of mitochondrial DNA expression

• Authors: Janusz Piechota , Roman Szczęsny , Kamila Wolanin , Aleksander Chlebowski , Ewa Bartnik
• Languages of publication: EN

Abstracts

EN

The influence of mutations in the mitochondrial DNA (mtDNA) on the bioenergetic metabolism of the cell is still poorly understood. Many of the mutations in the mtDNA affect the expression of the mitochondrial genome. Investigations on cells from patients are not easy, especially as the mitochondrial DNA is heteroplasmic and this state is changed in culture. Moreover, the nuclear background and the mitochondrial haplotype may affect the behaviour of cells. Transfer of patient mitochondria to rho zero cell lines is also not optimal as these cells in general have many nuclear changes which may also affect cell behaviour. Thus, we decided to use inhibitors of mitochondrial genome expression, such as thiamphenicol, ethidium bromide and dideoxycytidine to investigate the bioenergetic metabolism of HeLa cells. We found that oxidative phosphorylation and glycolysis participate equally in ATP production in HeLa cells and that decreased activity of the respiratory chain leads to increased glycolysis and the reduction of cell growth. Insufficient ATP production in the oxidative phosphorylation process was not compensated by increased proliferation of the mitochondria. However, we were able to show that there are some mechanisms compensating limited expression of the mitochondrial genome within the mitochondria. Experiments with dideoxycytidine revealed that 10-fold decrease of the mtDNA copy number resulted in almost normal activity of cytochrome c oxidase. We found that mtDNA depletion is compensated mostly on the level of RNA metabolism in the mitochondria. Thus, our results are in agreement with the hypothesis that transcription initiation rather than mtDNA copy number is a rate limiting factor for expression of the mitochondrial genome.

Keywords

EN

thiamphenicol; mtDNA depletion; HeLa; dideoxycytidine; mitochondrial biogenesis; ethidium bromide

• Publisher: Polish Biochemical Society
• Journal: Acta Biochimica Polonica
• Year: 2006
• Volume: 53
• Issue: 3
• Pages: 485-495

Dates

published

2006

received

2006-06-08

accepted

2006-08-14

revised

2006-08-14

(unknown)

2006-09-02

Contributors

author

Janusz Piechota

• Department of Genetics and Biotechnology, Warsaw University, Warszawa, Poland

author

Roman Szczęsny

• Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warszawa, Poland

author

Kamila Wolanin

• Department of Genetics and Biotechnology, Warsaw University, Warszawa, Poland

author

Aleksander Chlebowski

• Department of Genetics and Biotechnology, Warsaw University, Warszawa, Poland

author

Ewa Bartnik

• Department of Genetics and Biotechnology, Warsaw University, Warszawa, Poland

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