Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Av. da República (EAN) 2780-157 Oeiras, Portugal.
Eco-Bio, Instituto de Investigação Científica Tropical, Av. da República (EAN) 2784-505 Oeiras, Portugal.
Microbiology (Reading). 2012 Sep;158(Pt 9):2408-2418. doi: 10.1099/mic.0.056531-0. Epub 2012 Jun 14.
In spite of the large number of reports on the aerobic respiratory chain of Escherichia coli, from gene transcription regulation to enzyme kinetics and structural studies, an integrative perspective of this pathway is yet to be produced. Here, a multi-level analysis of the aerobic respiratory chain of E. coli was performed to find correlations between gene transcription, enzyme activity, growth dynamics, and supercomplex formation and composition. The transcription level of all genes encoding the aerobic respiratory chain of E. coli varied significantly in response to bacterial growth. Coordinated expression patterns were observed between the genes encoding NADH : quinone oxidoreductase and complex I (NDH-1), alternative NADH : quinone oxidoreductase (NDH-2) and cytochrome bdI, and also between sdhA and appC, encoding succinate dehydrogenase and cytochrome bdII, respectively. In general, the rates of the respiratory chain activities increased from mid-exponential to late-stationary phase, with no significant further variation occurring until the mid-stationary phase. Multi-level correlations between gene transcription, enzyme activity and growth dynamics were also found in this study. The previously reported NADH dehydrogenase and formate : oxygen oxidoreductase supercomplexes of E. coli were already assembled at mid-exponential phase and remained throughout growth. A new succinate oxidase supercomplex composed of succinate dehydrogenase and cytochrome bdII was identified, in agreement with the suggestion provided by the coordinated transcription of sdhA and appC.
尽管已经有大量关于大肠杆菌需氧呼吸链的报告,涵盖了从基因转录调控到酶动力学和结构研究等多个方面,但该途径仍缺乏综合视角。在此,我们对大肠杆菌的需氧呼吸链进行了多层次分析,以寻找基因转录、酶活性、生长动态以及超复合体形成和组成之间的相关性。大肠杆菌需氧呼吸链的所有基因转录水平在细菌生长过程中均发生显著变化。编码 NADH:醌氧化还原酶和复合物 I(NDH-1)、替代 NADH:醌氧化还原酶(NDH-2)和细胞色素 bdI 的基因之间以及编码琥珀酸脱氢酶和细胞色素 bdII 的 sdhA 和 appC 基因之间观察到协调表达模式。一般来说,呼吸链活性的速率从中期指数增长阶段增加到晚期稳定阶段,直到中期稳定阶段才会发生明显的进一步变化。本研究还发现了基因转录、酶活性和生长动态之间的多层次相关性。先前报道的大肠杆菌 NADH 脱氢酶和甲酸盐:氧氧化还原酶超复合体已经在中期指数增长阶段组装,并在整个生长过程中保持不变。鉴定出一种由琥珀酸脱氢酶和细胞色素 bdII 组成的新的琥珀酸氧化酶超复合体,这与 sdhA 和 appC 的协调转录所提出的建议一致。