ArticleApplied and environmental microbiology2003
Mutational analysis of the critical bases involved in activation of the AreR-regulated sigma54-dependent promoter in Acinetobacter sp. strain ADP1.
Article in Applied and environmental microbiology, 2003. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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15 citing papers in PubMed.
- Rewiring Aromatic Compound Consumption: Chromosomal Amplification and Evolution of a Foreign Pathway inACS synthetic biology · 2025Article
- Regulation of tricarboxylate transport and metabolism inApplied and environmental microbiology · 2024Article
- Regulation of l- and d-Aspartate Transport and Metabolism in Acinetobacter baylyi ADP1.Applied and environmental microbiology · 2022Article
- Development of a genetic toolset for the highly engineerable and metabolically versatile Acinetobacter baylyi ADP1.Nucleic acids research · 2020Article
- Rapid and assured genetic engineering methods applied to Acinetobacter baylyi ADP1 genome streamlining.Nucleic acids research · 2020Article
- Removal of aromatic inhibitors produced from lignocellulosic hydrolysates byBiotechnology for biofuels · 2019Article
- Functional Redundancy in the Hydroxycinnamate Catabolism Pathways of the Salt Marsh Bacterium Sagittula stellata E-37.Applied and environmental microbiology · 2018Article
- Vibrio fischeri DarR Directs Responses to d-Aspartate and Represents a Group of Similar LysR-Type Transcriptional Regulators.Journal of bacteriology · 2018Article
- Malonate degradation inMicrobiology (Reading, England) · 2017Article
- Article
- Deletion mutations caused by DNA strand slippage in Acinetobacter baylyi.Applied and environmental microbiology · 2006Article
- When coupled to natural transformation in Acinetobacter sp. strain ADP1, PCR mutagenesis is made less random by mismatch repair.Applied and environmental microbiology · 2005Article
- Toxicity caused by hydroxycinnamoyl-coenzyme A thioester accumulation in mutants of Acinetobacter sp. strain ADP1.Applied and environmental microbiology · 2004Article
- Acinetobacter sp. ADP1: an ideal model organism for genetic analysis and genome engineering.Nucleic acids research · 2004Article
- Hydroxycinnamate (hca) catabolic genes from Acinetobacter sp. strain ADP1 are repressed by HcaR and are induced by hydroxycinnamoyl-coenzyme A thioesters.Applied and environmental microbiology · 2003Article
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2 authors.
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Abstract
The areR gene in Acinetobacter sp. strain ADP1 regulates the expression of the areCBA genes, which determine growth on benzyl alkanoates. AreR is a member of the NtrC/XylR family of regulatory proteins as determined by sequence homology. Seventy-nine bases upstream of the start of transcription is a region carrying two overlapping inverted repeat (IR) sequences that we predict to be the AreR binding site, also known as the upstream activator site (UAS). IR1 is a near-perfect (16 of 17 bp) repeat separated by 1 bp, and IR2 consists of 9- and 7-bp perfect repeats with a 3-bp gap, with the central bases of the two arms of the repeat separated by 44 and 22 bp. We report here a method for site-directed mutagenesis of chromosomal genes in ADP1 in which linear fragments generated by overlap extension PCR are used to transform ADP1 via its natural transformation system and recombinants are selected by a marker exchange-eviction strategy with a newly created sacB-Km cassette. This method was used to generate 38 strains with designed mutations in the putative UAS upstream of areCBA. The effects of the mutations on areCBA expression were measured by enzyme assays of benzyl alcohol dehydrogenase (AreB) and by reporter gene assays of lacZ inserted into areA. Substitutions or deletions in IR1 had more deleterious effects upon expression when they were in its central region, which overlaps the left arm of IR2, than when they were in its outer regions. By contrast, substitutions in the right arm of IR2 resulted in mutants with relatively high expression levels compared to that of the wild type. Effects of deletions in the right arm of IR2 were very dependent upon the length of the deletion, with 3- or 5-bp deletions reducing expression by >90% whereas an 11-bp deletion in the same area reduced the expression levels by only 50%, suggesting that alterations in the distance and the orientation of the UAS relative to the -24, -12 sigma(54) promoter are critical.
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