Regulation of the Expression of nucS, a Key Component of the Mismatch Repair System in Mycobacteria

Mismatch repair (MMR) system alterations can trigger transient hypermutation, promoting adaptive mutations under stress, such as antibiotic exposure. While most organisms use MutS and MutL protein families for MMR, many archaea and actinobacteria, including the major human pathogen <i>Mycobacterium tuberculosis</i>, lack these components and instead rely on NucS, a structurally distinct enzyme driving a non-canonical MMR pathway. Given the role of MMR in mutation control, understanding how <i>nucS</i> expression is regulated could be essential for uncovering the molecular basis of antibiotic resistance development in mycobacteria. In this study, we characterized the <i>nucS</i> promoter and transcription start site in <i>Mycobacterium smegmatis</i>. We found that <i>nucS</i> expression declines during the stationary phase in both <i>M. smegmatis</i> and <i>M. tuberculosis</i>, paralleling replication activity and canonical MMR downregulation. Our data suggest that the alternative sigma factor σ^B may negatively regulate <i>nucS</i> expression during this phase. Additionally, we identified candidate compounds that may modulate <i>nucS</i> expression, underscoring its responsiveness to environmental cues. These findings enhance our understanding of mycobacterial stress responses and lay the groundwork for exploring antibiotic resistance mechanisms. Strikingly, our work reveals a case of double convergent evolution: both canonical (MutS/MutL) and non-canonical (NucS) pathways have independently evolved not only the same DNA repair function, but also similar regulatory frameworks for genome integrity preservation under stress conditions.