1887

Abstract

The () gene of ATCC 13032 encodes a regulatory protein belonging to the MocR subfamily of GntR-type transcription regulators and consisting of an amino-terminal winged helix–turn–helix DNA-binding domain and a carboxy-terminal aminotransferase-like domain. A defined deletion in the gene resulted in the decreased expression of the divergently orientated genes coding for the subunits of pyridoxal 5′-phosphate synthase. The mutant NJ0898 and the mutant AMH17 showed vitamin B auxotrophy that was restored by supplementing the growth medium with either pyridoxal, pyridoxal 5′-phosphate or pyridoxamine. The genetic organization of the 89 bp intergenic region was elucidated by mapping the 5′ ends of the respective transcripts, followed by detection of typical promoter sequences. Bioinformatic pattern searches and comparative genomics revealed three DNA motifs with the consensus sequence AAAGTGGW(−/T)CTA, overlapping the deduced promoter sequences and serving as candidate DNA-binding sites for PdxR. DNA band shift assays with the purified PdxR protein demonstrated the specific binding of the transcription regulator to double-stranded 40-mer sequences containing the detected motifs, thereby confirming the direct regulatory role of PdxR in activating the expression of the genes.

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2011-01-01
2024-03-29
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