, a pathway-specific transcriptional regulatory gene, positively controls polyoxin biosynthesis in subsp. Free

Abstract

The polyoxin (POL) biosynthetic gene cluster () was recently cloned from subsp. . A 3.3 kb DNA fragment carrying an obvious open reading frame (), whose deduced product shows sequence similarity to SanG of and PimR of , was revealed within the gene cluster. Disruption of abolished POL production, which could be complemented by the integration of a single copy of into the chromosome of the non-producing mutant. The introduction of an extra copy of in the wild-type strain resulted in increased production of POLs. The transcription start point (tsp) of was determined by S1 mapping. Reverse transcriptase PCR experiments showed that PolR is required for the transcription of 18 structural genes in the gene cluster. Furthermore, we showed that and , the respective first genes of two putative operons ( and ) consisting of 16 and 2 of these 18 genes, have similar promoter structures. Gel retardation assays indicated that PolR has specific DNA-binding activity for the promoter regions of and . Our data suggest that PolR acts in a positive manner to regulate POL production by activating the transcription of at least two putative operons in the gene cluster.

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2009-06-01
2024-03-29
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