Dukan S, Nyström T
Department of Cell and Molecular Biology-Microbiology, Göteborg University, Box 462, 405 30 Göteborg, Sweden.
J Biol Chem. 1999 Sep 10;274(37):26027-32. doi: 10.1074/jbc.274.37.26027.
Analysis of protein carbonylation demonstrates that the stasis-induced catalases and cytoplasmic superoxide dismutases (SOD) have a role in preventing accelerated protein oxidation during growth arrest of Escherichia coli cells. A larger number of proteins are carbonylated in cells lacking cytoplasmic SOD compared with cells lacking catalases, OxyR, or RpoS which, in turn, exhibit a larger number of oxidized proteins than the wild-type parent. Proteins exclusively oxidized during stasis in mutants lacking cytoplasmic SOD include GroEL, EF-G, and the acidic isoform of H-NS indicating that these mutants experience problems in peptide elongation and maintaining protein and DNA architecture. These mutants also survive stasis poorly. Likewise, but to a much lesser extent, mutations in oxyR, an oxidative stress regulator, shorten the life-span of stationary phase cells. The low plating efficiency of cells lacking OxyR is the result of their inability to grow on standard culture plates unless plating is performed anaerobically or with high concentration of catalase. In contrast, cells lacking cytoplasmic SOD appear to die prior to plating. Our data points to the importance of oxidative stress defense in stasis survival, and we also demonstrate that the life-span of growth-arrested wild-type E. coli cells can be significantly extended by omitting oxygen.
蛋白质羰基化分析表明,停滞诱导的过氧化氢酶和细胞质超氧化物歧化酶(SOD)在防止大肠杆菌细胞生长停滞期间蛋白质氧化加速方面发挥作用。与缺乏过氧化氢酶、OxyR或RpoS的细胞相比,缺乏细胞质SOD的细胞中有更多蛋白质发生羰基化,而缺乏过氧化氢酶、OxyR或RpoS的细胞又比野生型亲本表现出更多的氧化蛋白质。在缺乏细胞质SOD的突变体中,停滞期间专门被氧化的蛋白质包括GroEL、EF-G和H-NS的酸性同工型,这表明这些突变体在肽链延伸以及维持蛋白质和DNA结构方面存在问题。这些突变体在停滞期的存活能力也很差。同样地,但程度要小得多,氧化应激调节因子OxyR的突变会缩短稳定期细胞的寿命。缺乏OxyR的细胞平板接种效率低是因为它们无法在标准培养平板上生长,除非在厌氧条件下或使用高浓度过氧化氢酶进行接种。相比之下,缺乏细胞质SOD的细胞在接种前似乎就已死亡。我们的数据表明氧化应激防御在停滞期存活中的重要性,并且我们还证明,通过去除氧气,生长停滞的野生型大肠杆菌细胞的寿命可以显著延长。