Identification of the Mycobacterium tuberculosis GlnE promoter and its response to nitrogen availability

Adenylyltransferase, GlnE, has a predicted role in controlling the enzymic activity of glutamine synthetase, the key enzyme in ammonia assimilation. It was previously demonstrated that glnE is an essential gene in Mycobacterium tuberculosis. glnE is located downstream of glnA2, one of four glutamine synthetases. The expression of GlnE under various conditions was determined. Although a co-transcript of glnA2 and glnE was detectable, the major transcript was monocistronic. A transcriptional start site immediately upstream of glnE was identified and it was shown by site-directed mutagenesis that the predicted -10 region is a functional promoter. It was demonstrated that in a Mycobacterium smegmatis background M. tuberculosis P_glnE was up-regulated in ammonia- or glutamine-containing media.