Role of mitochondrial NADH kinase and NADPH supply in the respiratory chain activity of Saccharomyces cerevisiae

摘要

In Saccharomyces cerevisiae, the mitochondrial nicotinamide adenine dinucleotide hydride kinase Pos5p is required for a variety of essential cellular pathways, most importantly respiration. The Pos5p knockout strain pos5Δ grows poorly in non-fermentable media. A potential relationship between this respiratory deficiency and the ability of the cells to supply nicotinamide adenine dinucleotide phosphate (NADPH) was examined by analyzing the respiratory chain activity of pos5Δ and two NADP+-specific dehydrogenase mutants, idp1Δ and zwf1Δ. All of the respiratory chain complexes of pos5Δ exhibited poor relative activity of <26% at the middle-log phase and 62% at the stationary phase. The respiratory chain activity levels of idp1Δ and zwf1Δ also reduced to 22%–37% and 28%–84% at the middle-log phase, and 73%–81% and 67%–88% at the stationary phase, not as robustly as those of pos5Δ. The double-mutant idp1pos5Δ exhibited even lower activities of <20% at the middle-log phase, but zwf1pos5Δ showed similar activities with pos5Δ. The complemented strain POS5/pos5Δ exhibited 1.05- to 3-fold higher activities than pos5Δ. These data showed that Pos5p contributes to the maintenance of respiratory chain complex activities, with other NADPH sources, such as Idp1p and Zwf1p, making a smaller contribution. These contributions were partly related to the ability of the cells to supply NADPH, especially in the mitochondria.