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PMID:10071223
| Citation |
Pellicer, MT, Lynch, AS, De Wulf, P, Boyd, D, Aguilar, J and Lin, EC (1999) A mutational study of the ArcA-P binding sequences in the aldA promoter of Escherichia coli. Mol. Gen. Genet. 261:170-6 |
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| Abstract |
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. |
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| Keywords |
Aldehyde Dehydrogenase/genetics; Aldehyde Dehydrogenase/metabolism; Bacterial Outer Membrane Proteins/genetics; Bacterial Outer Membrane Proteins/metabolism; Base Sequence; Binding Sites; DNA, Bacterial/genetics; DNA, Bacterial/metabolism; Deoxyribonuclease I/metabolism; Electrophoresis; Escherichia coli/genetics; Escherichia coli/metabolism; Escherichia coli Proteins; Gene Expression Regulation, Bacterial; Gene Expression Regulation, Enzymologic; Molecular Sequence Data; Mutation; Phosphorylation; Promoter Regions, Genetic; Protein Binding; Repressor Proteins; Transcription, Genetic |
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Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
| GO:0006355: regulation of transcription, DNA-dependent |
IDA: Inferred from Direct Assay: |
P |
Figure 2. EMSA of arcA and arcA-P binding to aldA promoter. This figure shows arcA binds to the aldA promoter but Table 2 is needed to show that this binding regulates transcription. Table 2. The arcA binding region of the aldA promoter was mutated. This mutation relieved the aldA promoter from arcA repression during anaerobic conditions.
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complete |
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See also
References
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