Abstract
Mycobacterium tuberculosis is a formidable pathogen capable of establishing persistent infections within macrophages. To survive and thrive within the host environment, it has evolved intricate regulatory networks, including a diverse array of transcription factors that enable adaptation to various stresses encountered within the host. However, the mechanisms by which transcription factors regulate biofilm formation in M. tuberculosis remain incompletely understood. This study aimed to investigate the role of serC, encoding phosphoserine aminotransferase, and its regulation by NapR, a transcription factor, in mycobacterial physiology. NapR regulates serC through directly binding to its promoter. Notably, the regulatory effect and corresponding phenotypes vary due to distinct binding affinities of NapR for the serC promoter in different mycobacterial species. In Mycobacterium smegmatis, NapRMsm positively regulates biofilm formation, growth on solid media, and the transition from microcolonies to microcolonies by activating serCMsm. In the BCG vaccine, on the contrary, NapRBCG represses serCBCG, thus negatively regulating colony size and alleviating the growth inhibition caused by high concentrations of serine. Furthermore, proteomic analysis suggested NapR serves as a global transcriptional regulator in BCG vaccine strains by simultaneously modulating four metabolic pathways. These findings underscore the complex and strain-specific regulatory mechanisms governing serine metabolism in mycobacteria and provide valuable insights into the interplay between metabolism, gene regulation, and bacterial physiology.