A mutational study of the ArcA-P binding sequences in the aldA promoter of Escherichia coli
| Autor: | M. Teresa Pellicer, A. S. Lynch, Edmund C. C. Lin, Juan Aguilar, P. De Wulf, Dana Boyd |
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| Rok vydání: | 1999 |
| Předmět: |
DNA
Bacterial Electrophoresis Transcription Genetic Mutant Molecular Sequence Data Biology medicine.disease_cause Gene Expression Regulation Enzymologic Genetics medicine Consensus sequence Escherichia coli Deoxyribonuclease I Binding site Phosphorylation Promoter Regions Genetic Molecular Biology Gene Binding Sites Base Sequence Escherichia coli Proteins Promoter Gene Expression Regulation Bacterial Aldehyde Dehydrogenase Molecular biology Repressor Proteins Response regulator Biochemistry Coding strand Mutation Bacterial Outer Membrane Proteins Protein Binding |
| Zdroj: | Moleculargeneral genetics : MGG. 261(1) |
| ISSN: | 0026-8925 |
| Popis: | The aldA gene (encoding aldehyde dehydrogenase) of Escherichia coli is anaerobically repressed by ArcA-P, the phosphorylated response regulator of the ArcB/A two-component signal transduction system. The promoter region of aldA contains two 10-bp sequences (5'-TGTTAATTAA-3') that perfectly match the proposed ArcA-P binding consensus (5'-[A/T]GTTAATTA[A/T]-3'). One consensus sequence is on the coding strand (-13 to -4 from the transcriptional start point), whereas the other is on the template strand (position -2 to -11). In this study we used the aldA promoter to test the validity of the proposed consensus sequence. DNase I protection experiments confirmed the 10-bp sequence to be a strong ArcA-P binding site. Alteration of the wild-type sequence from 5'-TGTTAATTAAC-3' to 5'-TCTTAATTAAG-3' or 5'-TATTAATTAAT-3' by site-directed mutagenesis markedly decreased the in vitro affinity of the promoter region for ArcA-P, and abolished the anaerobic repression of mutant att lambda::phi (aldA'-lacZ) transcriptional reporter constructs. Both the in vitro and in vivo results therefore support the proposed consensus sequence. |
| Databáze: | OpenAIRE |
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