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Mycobacterium tuberculosis  has the ability to survive inside macrophages under acid-nitrosative stress.  M. tuberculosis Rv1685c  and its ortholog in  M. smegmatis, MSMEG_3765 , are induced on exposure to acid-nitrosative stress. Both genes are annotated as TetR transcriptional regulators, a family of proteins that regulate a wide range of cellular activities, including multidrug resistance, carbon catabolism and virulence. Here, we demonstrate that  MSMEG_3765  is co-transcribed with the upstream genes  MSMEG_3762  and  MSMEG_3763 , encoding efflux pump components. RTq-PCR and GFP-reporter assays showed that the  MSMEG_3762/63/65  gene cluster, and the orthologous region in  M. tuberculosis  ( Rv1687c/86c/85c ), was up-regulated in a  MSMEG_3765  null mutant, suggesting that  MSMEG_3765  acts as a repressor, typical of this family of regulators. We further defined the  MSMEG_3765  regulon using genome-wide transcriptional profiling and used reporter assays to confirm that the  MSMEG_3762/63/65  promoter was induced under acid-nitrosative stress. A putative 36 bp regulatory motif was identified upstream of the gene clusters in both  M. smegmatis  and  M. tuberculosis  and purified recombinant MSMEG_3765 protein was found to bind to DNA fragments containing this motif from both  M. smegmatis  and  M. tuberculosis  upstream regulatory regions. These results suggest that the TetR repressor  MSMEG_3765/Rv1685c  controls expression of an efflux pump with an, as yet, undefined role in the mycobacterial response to acid-nitrosative stress.

A Novel TetR-Like Transcriptional Regulator Is Induced in Acid-Nitrosative Stress and Controls Expression of an Efflux Pump in Mycobacteria