Weidner U, Geier S, Ptock A, Friedrich T, Leif H, Weiss H
Institut für Biochemie, Heinrich-Heine-Universität Düsseldorf, Federal Republic of Germany.
J Mol Biol. 1993 Sep 5;233(1):109-22. doi: 10.1006/jmbi.1993.1488.
The gene locus nuo of the proton-translocating NADH: ubiquinone oxidoreductase in Escherichia coli was identified by means of a DNA probe made by the polymerase chain reaction. The primers used for the reaction were derived from consensus sequences of the NAD(H)-binding subunit of mitochondrial NADH: ubiquinone oxidoreductase and the NAD(+)-reducing hydrogenase of Alcaligenes eutrophus. The nuo locus lies between minutes 49.2 and 49.6 on the E. coli chromosome and contains a cluster of 14 genes. They are bordered upstream by a sequence resembling sigma 70-dependent promoters and downstream by a sequence resembling rho-independent terminators. All 14 proteins derived from the nuo-genes are related to subunits of mitochondrial NADH: ubiquinone oxidoreductase, among which all subunits presumed to bind the substrates and to harbour the redox groups are found, as well as all seven mitochondrially encoded subunits. The E. coli enzyme seems to represent the minimal form of a proton-translocating NADH: ubiquinone oxidoreductase. The gene order in the nuo locus is conserved in comparison with other bacterial genomes and the chloroplast genome of higher plants. To some extent, the gene order correlates with the topological arrangement of the encoded subunits. The conception of modular evolution of NADH: ubiquinone oxidoreductase is further supported by the arrangement of the nuo-genes.
利用聚合酶链反应制备的DNA探针,鉴定了大肠杆菌中质子转运型NADH:泛醌氧化还原酶的基因位点nuo。用于该反应的引物源自线粒体NADH:泛醌氧化还原酶的NAD(H)结合亚基和嗜碱假单胞菌的NAD(+)还原氢化酶的共有序列。nuo基因位点位于大肠杆菌染色体上49.2分钟至49.6分钟之间,包含一组14个基因。它们的上游边界是一个类似于依赖sigma 70的启动子的序列,下游边界是一个类似于不依赖rho的终止子的序列。nuo基因产生的所有14种蛋白质都与线粒体NADH:泛醌氧化还原酶的亚基相关,其中发现了所有假定结合底物并含有氧化还原基团的亚基,以及所有7个线粒体编码的亚基。大肠杆菌的这种酶似乎代表了质子转运型NADH:泛醌氧化还原酶的最小形式。与其他细菌基因组和高等植物的叶绿体基因组相比,nuo基因位点中的基因顺序是保守的。在某种程度上,基因顺序与所编码亚基的拓扑排列相关。nuo基因的排列进一步支持了NADH:泛醌氧化还原酶模块化进化的概念。