The Two-Component System PhoPR of Clostridium acetobutylicum Is Involved in Phosphate-Dependent Gene Regulation
| Autor: | Maren Mix, Tomas Fiedler, Uta Meyer, Michael O. Glocker, Stefan Mikkat, Ralf-Jörg Fischer, Hubert Bahl |
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| Rok vydání: | 2008 |
| Předmět: |
DNA
Bacterial Clostridium acetobutylicum Operon Molecular Sequence Data Electrophoretic Mobility Shift Assay Microbiology Phosphates Bacterial Proteins Gene Order Electrophoresis Gel Two-Dimensional Gene Regulation Electrophoretic mobility shift assay Amino Acid Sequence Phosphorylation Promoter Regions Genetic Molecular Biology Regulation of gene expression Base Sequence biology Gene Expression Profiling Autophosphorylation Promoter Gene Expression Regulation Bacterial biochemical phenomena metabolism and nutrition biology.organism_classification Molecular biology Two-component regulatory system Response regulator bacteria Transcription Initiation Site Sequence Alignment Protein Binding Signal Transduction |
| Zdroj: | Journal of Bacteriology. 190:6559-6567 |
| ISSN: | 1098-5530 0021-9193 |
| DOI: | 10.1128/jb.00574-08 |
| Popis: | 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 P i -limited chemostat cultures and in response to P i pulses. A basic transcription level under high-phosphate conditions was shown, and a significant increase in mRNA transcript levels was found when external P i concentrations dropped below 0.3 mM. In two-dimensional gel electrophoresis experiments, a 2.5-fold increase in PhoP was observed under P i -limiting growth conditions compared to growth with an excess of P i . 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 . |
| Databáze: | OpenAIRE |
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