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InAmycolatopsis mediterranei U32, genes responsible for nitrate assimilation formed one operon,nasACKBDEF , whose transcription is induced by the addition of nitrate. Here, we characterized GlnR as a direct transcriptional activator for thenas operon. The GlnR-protected DNA sequences in the promoter region of thenas operon were characterized by DNase I footprinting assay, the previously deducedStreptomyces coelicolor double 22-bp GlnR binding consensus sequences comprisinga1 ,b1 ,a2 , andb2 sites were identified, and the sites were then mutated individually to test their roles in both the binding of GlnRin vitro and the GlnR-mediated transcriptional activationin vivo . The results clearly showed that only three GlnR binding sites (a1 ,b1 , andb2 sites) were required by GlnR for its specific binding to thenas promoter region and efficient activation of the transcription of thenas operon in U32, while thea2 site seemed unnecessary.

Three of Four GlnR Binding Sites Are Essential for GlnR-Mediated Activation of Transcription of the Amycolatopsis mediterranei nas Operon