Chu Zhaoqing, Li Juntao, Eshaghi Majid, Karuturi R Krishna Murthy, Lin Kui, Liu Jianhua
Systems Biology, Genome Institute of Singapore, Singapore 138672, Singapore.
BMC Genomics. 2007 Sep 15;8:323. doi: 10.1186/1471-2164-8-323.
DNA polymerase gamma(Pol-gamma) 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-gamma is required for mtDNA maintenance in the petite-negative fission yeast Schizosaccharomyces pombe.
We show that disruption of the nuclear gene pog1+ that encodes Pol-gamma is sufficient to deplete mtDNA in S. pombe. Cells bearing pog1Delta allele require substantial growth periods to form petite colonies. Mitotracker assays indicate that pog1Delta cells are defective in mitochondrial function and EM analyses suggest that pog1Delta cells lack normal mitochondrial structures. Depletion of mtDNA in pog1Delta cells is evident from quantitative real-time PCR assays. Genome-wide expression profiles of pog1Delta 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.
By showing that Pol-gamma 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.
已证明DNA聚合酶γ(Pol-γ)对于小菌落阳性出芽酵母酿酒酵母中线粒体基因组(mtDNA)的维持至关重要。缺乏线粒体的出芽酵母细胞表现出生长缓慢或小菌落表型。小菌落菌株无法在非发酵碳源上生长。然而,尚不清楚在小菌落阴性裂殖酵母粟酒裂殖酵母中mtDNA的维持是否需要Pol-γ。
我们表明,编码Pol-γ的核基因pog1+的破坏足以使粟酒裂殖酵母中的mtDNA耗尽。携带pog1Δ等位基因的细胞需要相当长的生长期才能形成小菌落。线粒体示踪分析表明pog1Δ细胞的线粒体功能存在缺陷,电子显微镜分析表明pog1Δ细胞缺乏正常的线粒体结构。定量实时PCR分析表明pog1Δ细胞中mtDNA明显减少。pog1Δ和其他无mtDNA细胞的全基因组表达谱显示,许多参与应激反应、有机化合物氧化产生能量、细胞碳水化合物代谢和能量储备代谢的基因被诱导表达。相反,许多编码参与氨基酸代谢和氧化磷酸化的蛋白质的基因被抑制。
通过证明Pol-γ对于mtDNA的维持至关重要且pog1+的破坏会改变全基因组表达谱,我们证明了在小菌落阴性粟酒裂殖酵母中,缺乏mtDNA的细胞表现出适应性核基因表达反应。