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Background  Bacteria use transcriptional regulation to respond to environmental stresses. Specifically, exposure to antibacterial drugs is deemed to be an atypical stress, and altering transcriptional regulation in response to such stress can increase bacterial drug resistance. However, only a few transcription factors that regulate drug resistance have been reported.    Results  In the present study, a GntR family transcription factor, encoded by the  MSMEG_0535 (Ms0535)  gene, was shown to be an isoniazid (INH) resistance regulator in  Mycobacterium smegmatis.  When the  Ms0535  gene was overexpressed, cells showed a significant increase in INH resistance. First, the interaction between Ms0535 and its own promoter was determined, and a conserved 26-bp palindromic DNA binding motif was identified using electrophoretic mobility shift and DNaseI footprinting assays. Second, quantitative reverse transcription-PCR assays showed that Ms0535 acted as a transcriptional activator, and positively regulated its own expression, as well as that of a permease encoded by the  MSMEG_0534 (Ms0534)  gene. Similar to the case for the  Ms0535  gene, a recombinant  Ms0534 -overexpressing strain also exhibited increased INH resistance compared with the wild-type strain. Furthermore, we showed that  Ms0535  and  Ms0534  deletion strains were more sensitive to INH than the wild-type strain. Interestingly, overexpressing  Ms0534  in the  Ms0535  deletion strain enhanced its INH resistance. In contrast, the  Ms0534  deletion strain was still sensitive to INH even when  Ms0535  was overexpressed. These findings suggest that Ms0534 is an effector protein that affects INH resistance in  M. smegmatis .    Conclusions  In summary, the GntR transcriptional regulator Ms0535 positively regulates INH resistance by transcriptionally regulating the expression of the Ms0534 permease in M. smegmatis. These results improve our understanding of the role of transcriptional regulation in INH drug resistance in mycobacteria.    Electronic supplementary material  The online version of this article (doi:10.1186/s12866-015-0556-8) contains supplementary material, which is available to authorized users.

A GntR family transcription factor positively regulates mycobacterial isoniazid resistance by controlling the expression of a putative permease