ArgR-independent induction and ArgR-dependent superinduction of the astCADBE operon in Escherichia coli
For Escherichia coli, growth in the absence of ammonia is termed nitrogen limited and results in the induction of genes that assimilate other nitrogen sources, a response mediated by sigma(54) and nitrogen regulator I (NR(I), also called NtrC). The astCADBE operon, which is required for growth with...
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| Vydáno v: | Journal of bacteriology Ročník 184; číslo 11; s. 2940 |
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| Hlavní autoři: | , |
| Médium: | Journal Article |
| Jazyk: | angličtina |
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01.06.2002
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| ISSN: | 0021-9193 |
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| Abstract | For Escherichia coli, growth in the absence of ammonia is termed nitrogen limited and results in the induction of genes that assimilate other nitrogen sources, a response mediated by sigma(54) and nitrogen regulator I (NR(I), also called NtrC). The astCADBE operon, which is required for growth with arginine as the sole nitrogen source, is moderately expressed during general nitrogen limitation and maximally expressed in the presence of arginine. The operon is also induced in stationary phase. Primer extension analysis of E. coli revealed the presence of a sigma(54)-dependent promoter utilized in exponential phase during nitrogen limitation and a sigma(S)-dependent promoter active during stationary phase. We used an ast-lacZ fusion to show that arginine stimulates expression, that ArgR, the arginine repressor, enhances expression from both promoters but is not essential, and that transcription by the two forms of the RNA polymerase is competitive and mutually exclusive. We demonstrated the binding of RNA polymerase holoenzymes, NR(I), and ArgR to the promoter region in vitro. We also reconstituted transcription from both promoters with purified components, which confirmed the accessory role of ArgR for the sigma(54)-dependent promoter. Thus, the ast operon exhibits nitrogen source-specific induction that is unique for an NR(I)-dependent gene. The transcriptional regulation of the ast operon in E. coli differs from that in Salmonella enterica serovar Typhimurium, in which ArgR is required for ast operon expression. |
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| AbstractList | For Escherichia coli, growth in the absence of ammonia is termed nitrogen limited and results in the induction of genes that assimilate other nitrogen sources, a response mediated by sigma(54) and nitrogen regulator I (NR(I), also called NtrC). The astCADBE operon, which is required for growth with arginine as the sole nitrogen source, is moderately expressed during general nitrogen limitation and maximally expressed in the presence of arginine. The operon is also induced in stationary phase. Primer extension analysis of E. coli revealed the presence of a sigma(54)-dependent promoter utilized in exponential phase during nitrogen limitation and a sigma(S)-dependent promoter active during stationary phase. We used an ast-lacZ fusion to show that arginine stimulates expression, that ArgR, the arginine repressor, enhances expression from both promoters but is not essential, and that transcription by the two forms of the RNA polymerase is competitive and mutually exclusive. We demonstrated the binding of RNA polymerase holoenzymes, NR(I), and ArgR to the promoter region in vitro. We also reconstituted transcription from both promoters with purified components, which confirmed the accessory role of ArgR for the sigma(54)-dependent promoter. Thus, the ast operon exhibits nitrogen source-specific induction that is unique for an NR(I)-dependent gene. The transcriptional regulation of the ast operon in E. coli differs from that in Salmonella enterica serovar Typhimurium, in which ArgR is required for ast operon expression.For Escherichia coli, growth in the absence of ammonia is termed nitrogen limited and results in the induction of genes that assimilate other nitrogen sources, a response mediated by sigma(54) and nitrogen regulator I (NR(I), also called NtrC). The astCADBE operon, which is required for growth with arginine as the sole nitrogen source, is moderately expressed during general nitrogen limitation and maximally expressed in the presence of arginine. The operon is also induced in stationary phase. Primer extension analysis of E. coli revealed the presence of a sigma(54)-dependent promoter utilized in exponential phase during nitrogen limitation and a sigma(S)-dependent promoter active during stationary phase. We used an ast-lacZ fusion to show that arginine stimulates expression, that ArgR, the arginine repressor, enhances expression from both promoters but is not essential, and that transcription by the two forms of the RNA polymerase is competitive and mutually exclusive. We demonstrated the binding of RNA polymerase holoenzymes, NR(I), and ArgR to the promoter region in vitro. We also reconstituted transcription from both promoters with purified components, which confirmed the accessory role of ArgR for the sigma(54)-dependent promoter. Thus, the ast operon exhibits nitrogen source-specific induction that is unique for an NR(I)-dependent gene. The transcriptional regulation of the ast operon in E. coli differs from that in Salmonella enterica serovar Typhimurium, in which ArgR is required for ast operon expression. For Escherichia coli, growth in the absence of ammonia is termed nitrogen limited and results in the induction of genes that assimilate other nitrogen sources, a response mediated by sigma(54) and nitrogen regulator I (NR(I), also called NtrC). The astCADBE operon, which is required for growth with arginine as the sole nitrogen source, is moderately expressed during general nitrogen limitation and maximally expressed in the presence of arginine. The operon is also induced in stationary phase. Primer extension analysis of E. coli revealed the presence of a sigma(54)-dependent promoter utilized in exponential phase during nitrogen limitation and a sigma(S)-dependent promoter active during stationary phase. We used an ast-lacZ fusion to show that arginine stimulates expression, that ArgR, the arginine repressor, enhances expression from both promoters but is not essential, and that transcription by the two forms of the RNA polymerase is competitive and mutually exclusive. We demonstrated the binding of RNA polymerase holoenzymes, NR(I), and ArgR to the promoter region in vitro. We also reconstituted transcription from both promoters with purified components, which confirmed the accessory role of ArgR for the sigma(54)-dependent promoter. Thus, the ast operon exhibits nitrogen source-specific induction that is unique for an NR(I)-dependent gene. The transcriptional regulation of the ast operon in E. coli differs from that in Salmonella enterica serovar Typhimurium, in which ArgR is required for ast operon expression. |
| Author | Kiupakis, Alexandros K Reitzer, Larry |
| Author_xml | – sequence: 1 givenname: Alexandros K surname: Kiupakis fullname: Kiupakis, Alexandros K organization: Department of Molecular and Cell Biology, The University of Texas at Dallas, Richardson, Texas 75083-0688, USA – sequence: 2 givenname: Larry surname: Reitzer fullname: Reitzer, Larry |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/12003934$$D View this record in MEDLINE/PubMed |
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| SubjectTerms | Acyltransferases - genetics Acyltransferases - metabolism Arginine Bacterial Proteins - genetics Bacterial Proteins - physiology Base Sequence Culture Media DNA Footprinting DNA-Binding Proteins - genetics DNA-Binding Proteins - metabolism DNA-Directed RNA Polymerases - genetics Escherichia coli - enzymology Escherichia coli - genetics Escherichia coli - growth & development Escherichia coli Proteins Gene Expression Regulation, Bacterial Molecular Sequence Data Nitrogen - metabolism Operon PII Nitrogen Regulatory Proteins Promoter Regions, Genetic Protein Binding Repressor Proteins - genetics Repressor Proteins - physiology RNA Polymerase Sigma 54 Sigma Factor - genetics Trans-Activators Transcription Factors Transcription, Genetic |
| Title | ArgR-independent induction and ArgR-dependent superinduction of the astCADBE operon in Escherichia coli |
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