Abstract

BackgroundDNA polymerase γ(Pol-γ) has been shown to be essential for maintenance of the mitochondrial genome (mtDNA) in the petite-positive budding yeast Saccharomyces cerevisiae. Budding yeast cells lacking mitochondria exhibit a slow-growing or petite-colony phenotype. Petite strains fail to grow on non-fermentable carbon sources. However, it is not clear whether the Pol-γ is required for mtDNA maintenance in the petite-negative fission yeast Schizosaccharomyces pombe.ResultsWe show that disruption of the nuclear gene pog1+ that encodes Pol-γ is sufficient to deplete mtDNA in S. pombe. Cells bearing pog1Δ allele require substantial growth periods to form petite colonies. Mitotracker assays indicate that pog1Δ cells are defective in mitochondrial function and EM analyses suggest that pog1Δ cells lack normal mitochondrial structures. Depletion of mtDNA in pog1Δ cells is evident from quantitative real-time PCR assays. Genome-wide expression profiles of pog1Δ and other mtDNA-less cells reveal that many genes involved in response to stimulus, energy derivation by oxidation of organic compounds, cellular carbohydrate metabolism, and energy reserve metabolism are induced. Conversely, many genes encoding proteins involved in amino acid metabolism and oxidative phosphorylation are repressed.ConclusionBy showing that Pol-γ is essential for mtDNA maintenance and disruption of pog1+ alters the genome-wide expression profiles, we demonstrated that cells lacking mtDNA exhibit adaptive nuclear gene expression responses in the petite-negative S. pombe.

Highlights

  • DNA polymerase γ(Pol-γ) has been shown to be essential for maintenance of the mitochondrial genome in the petite-positive budding yeast Saccharomyces cerevisiae

  • We show that the nuclear gene pog1+ is essential for mtDNA maintenance

  • BLAST analysis of the S. pombe Pol-γ revealed its homolog in various other species such as Homo sapiens, Rattus norvegicus, Mus musculus, Caenorhabditis elegans, Candida albicans, and S. cerevisiae

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Summary

Introduction

DNA polymerase γ(Pol-γ) has been shown to be essential for maintenance of the mitochondrial genome (mtDNA) in the petite-positive budding yeast Saccharomyces cerevisiae. Budding yeast cells lacking mitochondria exhibit a slow-growing or petite-colony phenotype. Petite strains fail to grow on non-fermentable carbon sources. It is not clear whether the Polγ is required for mtDNA maintenance in the petite-negative fission yeast Schizosaccharomyces pombe. Mitochondria are one of the organelles that contain its own genetic material, the mitochondrial genome (mtDNA). Cells depleted of mtDNA (ρ°) have been isolated through the treatment with ethidium bromide which results in the formation of tiny or petite colonies in S. cerevisiae [11]. The petite mutants fail to grow on media containing non-fermentable carbon sources owing to a lack of mitochondrial functions. The isolated petites exhibited severely retarded growth phenotypes that impeded further genetic analyses of these petite strains

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