Weerakoon Dilan R, Olson Jonathan W
Department of Microbiology, North Carolina State University, Raleigh, North Carolina 27695, USA.
J Bacteriol. 2008 Feb;190(3):915-25. doi: 10.1128/JB.01647-07. Epub 2007 Dec 7.
Campylobacter jejuni encodes 12 of the 14 subunits that make up the respiratory enzyme NADH:ubiquinone oxidoreductase (also called complex I). The two nuo genes not present in C. jejuni encode the NADH dehydrogenase, and in their place in the operon are the novel genes designated Cj1575c and Cj1574c. A series of mutants was generated in which each of the 12 nuo genes (homologues to known complex I subunits) was disrupted or deleted. Each of the nuo mutants will not grow in amino acid-based medium unless supplemented with an alternative respiratory substrate such as formate. Unlike the nuo genes, Cj1574c is an essential gene and could not be disrupted unless an intact copy of the gene was provided at an unrelated site on the chromosome. A nuo deletion mutant can efficiently respire formate but is deficient in alpha-ketoglutarate respiratory activity compared to the wild type. In C. jejuni, alpha-ketoglutarate respiration is mediated by the enzyme 2-oxoglutarate:acceptor oxidoreductase; mutagenesis of this enzyme abolishes alpha-ketoglutarate-dependent O2 uptake and fails to reduce the electron transport chain. The electron acceptor for 2-oxoglutarate:acceptor oxidoreductase was determined to be flavodoxin, which was also determined to be an essential protein in C. jejuni. A model is presented in which CJ1574 mediates electron flow into the respiratory transport chain from reduced flavodoxin and through complex I.
空肠弯曲菌编码构成呼吸酶NADH:泛醌氧化还原酶(也称为复合体I)的14个亚基中的12个。空肠弯曲菌中不存在的两个nuo基因编码NADH脱氢酶,在操纵子中的相应位置是新基因Cj1575c和Cj1574c。构建了一系列突变体,其中12个nuo基因(与已知复合体I亚基同源)中的每一个都被破坏或缺失。除非添加替代呼吸底物(如甲酸),否则每个nuo突变体都不能在氨基酸培养基中生长。与nuo基因不同,Cj1574c是一个必需基因,除非在染色体上不相关的位点提供该基因的完整拷贝,否则无法被破坏。一个nuo缺失突变体可以有效地利用甲酸进行呼吸,但与野生型相比,其α-酮戊二酸呼吸活性不足。在空肠弯曲菌中,α-酮戊二酸呼吸由2-氧代戊二酸:受体氧化还原酶介导;该酶的诱变消除了依赖α-酮戊二酸的氧气摄取,并且未能减少电子传递链。确定2-氧代戊二酸:受体氧化还原酶的电子受体为黄素氧还蛋白,该蛋白在空肠弯曲菌中也是必需蛋白。提出了一个模型,其中CJ1574介导电子从还原的黄素氧还蛋白流入呼吸传递链并通过复合体I。