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

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.