Camougrand N M, Mouassite M, Velours G M, Guérin M G
Institut de Biochimie et Génétique Cellulaires du CNRS, Université Victor Segalen Bordeaux 2, 1 rue Camille Saint-Saëns, Bordeaux, 33077, France.
Arch Biochem Biophys. 2000 Mar 1;375(1):154-60. doi: 10.1006/abbi.1999.1655.
Since it was shown in previous work that NCA3 (one of the four genes of the SUN family) is involved in mitochondrial protein synthesis regulation, the effect of the other members of this gene family was tested. UTH1 (but not SUN4 or SIM1) was also shown to interfere with mitochondria biogenesis. In Deltauth1 cells, cytochromes aa(3), c, and b were lowered by 25 and 15%, respectively. In the double-null mutant Deltauth1Deltanca3, only cytochrome aa(3) was lowered by 50% relative to the wild type. However, the ratio of cellular respiration to cytochrome oxidase was greatly enhanced in the double-null mutant. Measurements on whole lysed cells showed that another mitochondrial enzyme, citrate synthase, was also lowered in Deltauth1 and Deltauth1Deltanca3 whereas hexokinase was not. Electron micrographs showed no difference in global mitochondria content in Deltauth1Deltanca3, but mitochondria appeared less dense to electrons compared to the wild type. Cardiolipin and mtDNA were equivalent in parental and mutant strains. Measurements on isolated mitochondria showed that the cyt aa(3)/cyt b ratio was also lowered in Deltauth1Deltanca3, but the control exerted by the oxidase on the respiratory flux was higher. The activity of other mitochondrial complexes versus oxidase was equivalent in mutants compared to the wild type. These results suggest that the protein equipment could be lowered in mitochondria from strains inactivated for UTH1.
由于在之前的研究中已表明NCA3(SUN家族四个基因之一)参与线粒体蛋白质合成调控,因此对该基因家族的其他成员的作用进行了测试。结果显示UTH1(而非SUN4或SIM1)也会干扰线粒体生物发生。在Δuth1细胞中,细胞色素aa(3)、c和b分别降低了25%和15%。在双缺失突变体Δuth1Δca3中,相对于野生型,只有细胞色素aa(3)降低了50%。然而,双缺失突变体中的细胞呼吸与细胞色素氧化酶的比率大大提高。对全裂解细胞的测量表明,另一种线粒体酶柠檬酸合酶在Δuth1和Δuth1Δca3中也有所降低,而己糖激酶则没有。电子显微镜照片显示,Δuth1Δca3中的整体线粒体含量没有差异,但与野生型相比,线粒体对电子的密度似乎较低。亲代菌株和突变体菌株中的心磷脂和线粒体DNA相当。对分离线粒体的测量表明,Δuth1Δca3中的细胞色素aa(3)/细胞色素b比率也降低了,但氧化酶对呼吸通量的控制更高。与野生型相比,突变体中其他线粒体复合物与氧化酶的活性相当。这些结果表明,对于UTH1失活的菌株,其线粒体中的蛋白质成分可能会减少。