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Abstract

Northern blot analysis and a GFP-based reporter assay showed that , which encodes glucose-6-phosphate dehydrogenase, was highly induced when KT2440 was cultured in minimal medium containing glucose or gluconate. However, expression was not detected in the presence of pyruvate or succinate. The use of a knockout mutant of HexR, a putative transcription regulator, resulted in constitutively high expression of , regardless of the carbon source. An electrophoretic mobility shift assay showed that HexR protein binds to the promoter region and that HexR binding is inhibited by 2-keto-3-deoxy-6-phosphogluconate (KDPG). Despite the presence of gluconate, the mutant (non-KDPG producer) was not able to induce the gene. The mutant (KDPG overproducer) featured a constitutively high level of expression. Interestingly, was also highly expressed in the presence of oxidative stress-inducing reagents. The level of induction in wild-type cells undergoing oxidative stress did not differ significantly from that observed with the mutant under normal conditions. Interestingly, the mutant was more tolerant of oxidative stress than the wild-type. Expression of was induced by oxidative stress in the mutant. Thus, KDPG, a real inducer of gene expression, was not necessary for oxidative-stress induction. binding of HexR to its cognate promoter region was diminished by menadione and cumene hydroperoxide. The data suggested that HexR might be a dual-sensing regulator of induction that is able to respond to both KDPG and oxidative stress.

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2008-12-01
2020-04-09
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