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Hydrogen peroxide (H 2 O 2 ) is a natural oxidant produced by aerobic organisms and gives rise to oxidative damage, including DNA mutations, protein inactivation and lipid damage. The genus  Mycobacterium  utilizes redox sensors and H 2 O 2  scavenging enzymes for the detoxification of H 2 O 2 . To date, the precise response to oxidative stress has not been fully elucidated. Here, we compared the effects of different levels of H 2 O 2  on transcription in  M .  smegmatis  using RNA-sequencing. A 0.2 mM H 2 O 2  treatment had little effect on the growth and viability of  M .  smegmatis  whereas 7 mM H 2 O 2  was lethal. Analysis of global transcription showed that 0.2 mM H 2 O 2  induced relatively few changes in gene expression, whereas a large proportion of the mycobacterial genome was found to be differentially expressed after treatment with 7 mM H 2 O 2 . Genes differentially expressed following treatment with 0.2 mM H 2 O 2  included those coding for proteins involved in glycolysis-gluconeogenesis and fatty acid metabolism pathways, and expression of most genes encoding ribosomal proteins was lower following treatment with 7 mM H 2 O 2 . Our analysis shows that  M .  smegmatis  utilizes the sigma factor MSMEG_5214 in response to 0.2 mM H 2 O 2 , and the RpoE1 sigma factors MSMEG_0573 and MSMEG_0574 in response to 7 mM H 2 O 2 . In addition, different transcriptional regulators responded to different levels of H 2 O 2 : MSMEG_1919 was induced by 0.2 mM H 2 O 2 , while high-level induction of DevR occurred in response to 7 mM H 2 O 2 . We detected the induction of different detoxifying enzymes, including genes encoding KatG, AhpD, TrxB and Trx, at different levels of H 2 O 2  and the detoxifying enzymes were expressed at different levels of H 2 O 2 . In conclusion, our study reveals the changes in transcription that are induced in response to different levels of H 2 O 2  in  M .  smegmatis .

Distinct Responses of Mycobacterium smegmatis to Exposure to Low and High Levels of Hydrogen Peroxide