Cotter P A, Chepuri V, Gennis R B, Gunsalus R P
Department of Microbiology and Molecular Genetics, University of California, Los Angeles 90024-1489.
J Bacteriol. 1990 Nov;172(11):6333-8. doi: 10.1128/jb.172.11.6333-6338.1990.
The aerobic respiratory chain of Escherichia coli contains two terminal oxidases that catalyze the oxidation of ubiquinol-8 and the reduction of oxygen to water. They are the cytochrome o oxidase complex encoded by cyoABCDE and the cytochrome d oxidase complex encoded by cydAB. To determine how these genes are regulated in response to a variety of environmental stimuli, including oxygen, we examined their expression by using lacZ protein fusions in wild-type and fnr mutant strains of E. coli. Anaerobic growth resulted in a 140-fold repression of cyoA'-'lacZ expression relative to aerobic growth and a 3-fold increase in cydA'-'lacZ expression. Anaerobic repression of both fusions was mediated in part by the fnr gene product, as evidenced by a 30-fold derepression of cyoA'-'lacZ expression and a 4-fold derepression of cydA'-'lacZ expression in an fnr deletion strain. Supplying wild-type fnr in trans restored wild-type repression for both fusions. Fnr thus functions as an anaerobic repressor of both cyoABCDE and cydAB expression. Reduced-minus-oxidized difference spectrum analyses of cell membranes confirmed the effect of the fnr gene product on the production of cytochrome d oxidase in the cell. Based on the pattern of anaerobic cydAB expression observed, we propose the existence of a second, as yet unidentified, regulatory element that must function either to activate cydAB expression as oxygen becomes limiting or to repress cydAB expression aerobically. Whereas cytochrome o oxidase encoded by cyoABCDE appears to be produced only under oxygen-rich growth conditions, in keeping with its biochemical properties, cytochrome d oxidase is expressed moderately aerobically and is elevated yet further when oxygen becomes limiting so that the organism can cope better under oxygen starvation conditions. We also examined cyoABCDE and cydAB expression in response to growth on alternative carbon compounds and to changes in the culture medium pH and osmolarity.
大肠杆菌的需氧呼吸链包含两种末端氧化酶,它们催化泛醌-8的氧化以及氧还原为水的过程。这两种酶分别是由cyoABCDE编码的细胞色素o氧化酶复合物和由cydAB编码的细胞色素d氧化酶复合物。为了确定这些基因如何响应包括氧气在内的各种环境刺激进行调控,我们利用大肠杆菌野生型和fnr突变株中的lacZ蛋白融合来检测它们的表达情况。与有氧生长相比,厌氧生长导致cyoA'-'lacZ表达受到140倍的抑制,而cydA'-'lacZ表达增加了3倍。两种融合蛋白的厌氧抑制部分是由fnr基因产物介导的,这一点在fnr缺失菌株中cyoA'-'lacZ表达30倍的去抑制和cydA'-'lacZ表达4倍的去抑制中得到了证明。反式提供野生型fnr可恢复两种融合蛋白的野生型抑制作用。因此,Fnr作为cyoABCDE和cydAB表达的厌氧阻遏物发挥作用。细胞膜的还原态减去氧化态差光谱分析证实了fnr基因产物对细胞中细胞色素d氧化酶产生的影响。基于观察到的厌氧cydAB表达模式,我们提出存在第二种尚未确定的调控元件,它必定在氧气变得有限时激活cydAB表达,或者在有氧时抑制cydAB表达。与cyoABCDE编码的细胞色素o氧化酶的生化特性一致,它似乎仅在富氧生长条件下产生,而细胞色素d氧化酶在有氧时适度表达,当氧气变得有限时表达进一步升高,以便生物体在缺氧条件下能更好地应对。我们还检测了cyoABCDE和cydAB在利用替代碳源生长以及培养基pH和渗透压变化时的表达情况。