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首页> 外文期刊>Journal of bacteriology >The Two-Component System PhoPR of Clostridium acetobutylicum Is Involved in Phosphate-Dependent Gene Regulation
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The Two-Component System PhoPR of Clostridium acetobutylicum Is Involved in Phosphate-Dependent Gene Regulation

机译：丙酮丁醇梭菌的两组分系统PhoPR参与了磷酸盐依赖性基因的调控。

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摘要

The phoPR gene locus of Clostridium acetobutylicum ATCC 824 comprises two genes, phoP and phoR. Deduced proteins are predicted to represent a response regulator and sensor kinase of a phosphate-dependent two-component regulatory system. We analyzed the expression patterns of phoPR in Pi-limited chemostat cultures and in response to Pi pulses. A basic transcription level under high-phosphate conditions was shown, and a significant increase in mRNA transcript levels was found when external Pi concentrations dropped below 0.3 mM. In two-dimensional gel electrophoresis experiments, a 2.5-fold increase in PhoP was observed under Pi-limiting growth conditions compared to growth with an excess of Pi. At least three different transcription start points for phoP were determined by primer extension analyses. Proteins PhoP and an N-terminally truncated *PhoR were individually expressed heterologously in Escherichia coli and purified. Autophosphorylation of *PhoR and phosphorylation of PhoP were shown in vitro. Electromobility shift assays proved that there was a specific binding of PhoP to the promoter region of the phosphate-regulated pst operon of C. acetobutylicum.
机译：丙酮丁醇梭菌（Clostridium acetobutylicum） ATCC 824的 phoPR 基因位点包含两个基因： phoP 和 phoR 。推导的蛋白质预计将代表一个依赖磷酸盐的两组分调节系统的响应调节器和传感器激酶。我们分析了 phoPR 在P i 限制的恒化器培养物中的表达模式，以及响应P i 脉冲的表达模式。显示了高磷酸盐条件下的基本转录水平，当外部P i 浓度降至0.3 mM以下时，mRNA转录水平显着增加。在二维凝胶电泳实验中，与限制P i 的生长相比，在限制P i 的生长条件下PhoP增加了2.5倍。通过引物延伸分析确定了 phoP 的至少三个不同的转录起始点。蛋白质PhoP和N末端截短的* PhoR在大肠杆菌中分别异源表达并纯化。体外显示* PhoR的自磷酸化和PhoP的磷酸化。电迁移率分析证明PhoP与磷酸调节的 C的 pst 操纵子的启动子区域存在特异性结合。乙草胺。

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