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Loss of mitochondrial DNA under genotoxic stress conditions in the absence of the yeast DNA helicase Pif1p occurs independently of the DNA helicase Rrm3p

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Abstract

How the cellular amount of mitochondrial DNA (mtDNA) is regulated under normal conditions and in the presence of genotoxic stress is less understood. We demonstrate that the inefficient mtDNA replication process of mutant yeast cells lacking the PIF1 DNA helicase is partly rescued in the absence of the DNA helicase RRM3. The rescue effect is likely due to the increase in the deoxynucleoside triphosphates (dNTPs) pool caused by the lack of RRM3. In contrast, the Pif1p-dependent mtDNA breakage in the presence and absence of genotoxic stress is not suppressed if RRM3 is lacking suggesting that this phenotype is likely independent of the dNTP pool. Pif1 protein (Pif1p) was found to stimulate the incorporation of dNTPs into newly synthesised mtDNA of gradient-purified mitochondria. We propose that Pif1p that acts likely as a DNA helicase in mitochondria affects mtDNA replication directly. Possible roles of Pif1p include the resolution of secondary DNA and/or DNA/RNA structures, the temporarily displacement of tightly bound mtDNA-binding proteins, or the stabilization of the mitochondrial replication complex during mtDNA replication.

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Acknowledgments

We thank Drs. Ronald Butow, Donna Gordon, Debkumar Pain, and Carolyn Suzuki for providing reagents. We are especially grateful to Drs. Donna Gordon, Chee-Gun Lee, Debkumar Pain, and Carolyn Suzuki for discussions and advice in cell biological aspects of mitochondria. This work is supported by a startup grant from the Department of Cell Biology and Molecular Medicine of the University of Medicine and Dentistry NJ, by a grant from the Foundation of the University of Medicine and Dentistry, NJ, and by a grant from the New Jersey Commission on Cancer Research (05-1975-CCR-EO).

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  1. Department of Cell Biology and Molecular Medicine, New Jersey Medical School, University of Medicine and Dentistry of New Jersey, 185 South Orange Avenue, Newark, NJ, 07101-1709, USA

    Xin Cheng, Yong Qin & Andreas S. Ivessa

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  1. Xin Cheng
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  2. Yong Qin
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  3. Andreas S. Ivessa
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Correspondence to Andreas S. Ivessa.

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Communicated by A. Aguilera.

X. Cheng, Y. Qin contributed equally to this work.

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Cheng, X., Qin, Y. & Ivessa, A.S. Loss of mitochondrial DNA under genotoxic stress conditions in the absence of the yeast DNA helicase Pif1p occurs independently of the DNA helicase Rrm3p. Mol Genet Genomics 281, 635–645 (2009). https://doi.org/10.1007/s00438-009-0438-6

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