Response of Mycobacterium smegmatis to the Cytochrome bcc Inhibitor Q203

For the design of next-generation tuberculosis chemotherapy, insight into bacterial defence against drugs is required. Currently, targeting respiration has attracted strong attention for combatting drug-resistant mycobacteria. Q203 (telacebec), an inhibitor of the cytochrome bcc complex in the mycobacterial respiratory chain, is currently evaluated in phase-2 clinical trials. Q203 has bacteriostatic activity against M. tuberculosis, which can be converted to bactericidal activity by concurrently inhibiting an alternative branch of the mycobacterial respiratory chain, cytochrome bd. In contrast, non-tuberculous mycobacteria, such as Mycobacterium smegmatis, show only very little sensitivity to Q203. In this report, we investigated factors that M. smegmatis employs to adapt to Q203 in the presence or absence of a functional cytochrome bd, especially regarding its terminal oxidases. In the presence of a functional cytochrome bd, M. smegmatis responds to Q203 by increasing the expression of cytochrome bcc as well as of cytochrome bd, whereas a M. smegmatis bd-KO strain adapted to Q203 by increasing the expression of cytochrome bcc. Interestingly, single-cell studies revealed cell-to-cell variability in drug adaptation. We also investigated the role of a putative second cytochrome bd isoform postulated for M. smegmatis. Although this putative isoform showed differential expression in response to Q203 in the M. smegmatis bd-KO strain, it did not display functional features similar to the characterised cytochrome bd variant.

Automated summary

Insight into bacterial defense mechanisms against drugs is crucial for the design of next-generation tuberculosis chemotherapy. This study investigates the adaptation strategies of Mycobacterium smegmatis to the drug Q203, particularly focusing on the role of cytochrome bd in the respiratory chain. The results show that M. smegmatis responds to Q203 by increasing the expression of cytochrome bcc and cytochrome bd in the presence of functional cytochrome bd. In the absence of functional cytochrome bd, M. smegmatis adapts to Q203 by increasing the expression of cytochrome bcc. Single-cell studies also reveal cell-to-cell variability in drug adaptation.