The regulatory role of Streptomyces coelicolor TamR in central metabolism

Trans -aconitate methyltransferase regulator (TamR) is a member of the ligand-responsive multiple antibiotic resistance regulator (MarR) family of transcription factors. In Streptomyces coelicolor , TamR regulates transcription of tamR (encoding TamR), tam (encoding trans -aconitate methyltransferase) and sacA (encoding aconitase); up-regulation of these genes promotes metabolic flux through the citric acid cycle. DNA binding by TamR is attenuated and transcriptional derepression is achieved on binding of ligands such as citrate and trans -aconitate to TamR. In the present study, we show that three additional genes are regulated by S. coelicolor TamR. Genes encoding malate synthase ( aceB1 ; SCO6243 ), malate dehydrogenase ( mdh ; SCO4827 ) and isocitrate dehydrogenase ( idh ; SCO7000 ) are up-regulated in vivo when citrate and trans -aconitate accumulate, and TamR binds the corresponding gene promoters in vitro , a DNA binding that is attenuated by cognate ligands. Mutations to the TamR binding site attenuate DNA binding in vitro and result in constitutive promoter activity in vivo . The predicted TamR binding sites are highly conserved in the promoters of these genes in Streptomyce s species that encode divergent tam–tamR gene pairs, suggesting evolutionary conservation. Like aconitase and trans -aconitate methyltransferase, malate dehydrogenase, isocitrate dehydrogenase and malate synthase are closely related to the citric acid cycle, either catalysing individual reaction steps or, in the case of malate synthase, participating in the glyoxylate cycle to produce malate that enters the citric acid cycle to replenish the intermediate pool. Taken together, our data suggest that TamR plays an important and conserved role in promoting metabolic flux through the citric acid cycle. Abbreviations: EGFP, enhanced green fluorescent protein; EMSA, electrophoretic mobility shift assay(s); MarR, multiple antibiotic resistance regulator; qRT-PCR, quantitative reverse transcription–PCR; TamR, trans-aconitate methyltransferase regulator; TBE, Tris/borate/EDTA