Gbelská Y, Subík J, Svoboda A, Goffeau A, Kovác L
Eur J Biochem. 1983 Feb 1;130(2):281-6. doi: 10.1111/j.1432-1033.1983.tb07148.x.
Cells of the yeast Saccharomyces cerevisiae could be depleted of their intramitochondrial ATP bu culturing on glucose in the presence of antimycin A, which prevents production of ATP in mitochondria, along with bongkrekic acid, which prevents transport of ATP from the cytosol into mitochondria. Alternatively, the depletion could be achieved by culturing respiration-deficient mutants in the presence of bongkrekic acid. The depleted cells of the respiration-deficient mutant did not grow on glucose in a synthetic medium and growth for a few generations was made possible by adding peptone, yeast extract or some amino acids into the medium. The depleted cells did not differ from control cells in their content of amino acids, proteins, nucleic acids and major phospholipids and had preserved the ability to carry on protein and nucleic acid syntheses and to mate to other cells. No conspicuous cytological differences were found between the control and depleted cells. After culturing in a semi-synthetic medium in the presence of bongkrekic acid the cells of the respiration-deficient mutant exhibited almost no cytochrome c in their spectra and their azide-sensitive ATPase activity was drastically reduced. The results suggest that intramitochondrial syntheses of some low-molecular compounds as well as import and/or assembly of some cytoplasmically synthesized mitochondrial proteins into mitochondria may be impaired in cells lacking intramitochondrial ATP and this may be responsible for their inability to grow and multiply.
在抗霉素A(可阻止线粒体中ATP的产生)和邦克雷酸(可阻止ATP从细胞质溶胶转运到线粒体)存在的情况下,通过在葡萄糖上培养,酿酒酵母细胞的线粒体内ATP可以被耗尽。或者,通过在邦克雷酸存在的情况下培养呼吸缺陷型突变体也可以实现耗尽。呼吸缺陷型突变体的耗尽细胞在合成培养基中不能在葡萄糖上生长,通过向培养基中添加蛋白胨、酵母提取物或一些氨基酸,使其能够生长几代。耗尽细胞在氨基酸、蛋白质、核酸和主要磷脂的含量上与对照细胞没有差异,并且保留了进行蛋白质和核酸合成以及与其他细胞交配的能力。对照细胞和耗尽细胞之间未发现明显的细胞学差异。在邦克雷酸存在的情况下,在半合成培养基中培养后,呼吸缺陷型突变体的细胞在光谱中几乎没有细胞色素c,并且其对叠氮化物敏感的ATP酶活性急剧降低。结果表明,在缺乏线粒体内ATP的细胞中,一些低分子化合物的线粒体内合成以及一些细胞质合成的线粒体蛋白向线粒体的导入和/或组装可能受损,这可能是它们无法生长和繁殖的原因。