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大肠杆菌K-12生长速率、EIIACrr磷酸化与细胞内环磷酸腺苷水平之间的相关性

Correlation between growth rates, EIIACrr phosphorylation, and intracellular cyclic AMP levels in Escherichia coli K-12.

作者信息

Bettenbrock Katja, Sauter Thomas, Jahreis Knut, Kremling Andreas, Lengeler Joseph W, Gilles Ernst-Dieter

机构信息

MPI für Dynamik Komplexer Technischer Systeme, Sandtorstr.1, 39106 Magdeburg, Germany.

出版信息

J Bacteriol. 2007 Oct;189(19):6891-900. doi: 10.1128/JB.00819-07. Epub 2007 Aug 3.

Abstract

In Escherichia coli K-12, components of the phosphoenolpyruvate-dependent phosphotransferase systems (PTSs) represent a signal transduction system involved in the global control of carbon catabolism through inducer exclusion mediated by phosphoenolpyruvate-dependent protein kinase enzyme IIA(Crr) (EIIA(Crr)) (= EIIA(Glc)) and catabolite repression mediated by the global regulator cyclic AMP (cAMP)-cAMP receptor protein (CRP). We measured in a systematic way the relation between cellular growth rates and the key parameters of catabolite repression, i.e., the phosphorylated EIIA(Crr) (EIIA(Crr) approximately P) level and the cAMP level, using in vitro and in vivo assays. Different growth rates were obtained by using either various carbon sources or by growing the cells with limited concentrations of glucose, sucrose, and mannitol in continuous bioreactor experiments. The ratio of EIIA(Crr) to EIIA(Crr) approximately P and the intracellular cAMP concentrations, deduced from the activity of a cAMP-CRP-dependent promoter, correlated well with specific growth rates between 0.3 h(-1) and 0.7 h(-1), corresponding to generation times of about 138 and 60 min, respectively. Below and above this range, these parameters were increasingly uncoupled from the growth rate, which perhaps indicates an increasing role executed by other global control systems, in particular the stringent-relaxed response system.

摘要

在大肠杆菌K-12中,磷酸烯醇丙酮酸依赖性磷酸转移酶系统(PTSs)的组分代表一种信号转导系统,该系统通过磷酸烯醇丙酮酸依赖性蛋白激酶IIA(Crr)(EIIA(Crr))(= EIIA(Glc))介导的诱导物排除以及全局调节因子环腺苷酸(cAMP)-cAMP受体蛋白(CRP)介导的分解代谢物阻遏,参与碳分解代谢的全局调控。我们通过体外和体内试验,系统地测量了细胞生长速率与分解代谢物阻遏关键参数之间的关系,即磷酸化的EIIA(Crr)(EIIA(Crr)P)水平和cAMP水平。通过使用各种碳源,或者在连续生物反应器实验中用有限浓度的葡萄糖、蔗糖和甘露醇培养细胞,获得了不同的生长速率。从cAMP-CRP依赖性启动子的活性推导得出的EIIA(Crr)与EIIA(Crr)P的比率以及细胞内cAMP浓度,与0.3 h⁻¹至0.7 h⁻¹之间的比生长速率具有良好的相关性,分别对应约138分钟和60分钟的代时。在此范围以下和以上,这些参数与生长速率的解偶联程度越来越高,这可能表明其他全局控制系统,特别是严谨-松弛反应系统发挥的作用越来越大。

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