Mycobacterium Tuberculosis Rv0341 Plays A Role In Bacterial Physiology And Antimicrobial Resistance Via Changing The Cell Wall Permeability

Tuberculosis(TB) is a major global health problem. It causes ill-health among millions of people each year and ranks alongside the human immunodeficiency virus(HIV) as a leading cause of death worldwide.Mycobacterium tuberculosis, is the causative agent of human tuberculosis, is most successful human pathogen that can survival and replicate within macrophages. Throughout infection, M. tuberculosis can sense the environmental subtle fluctuations and reprogram metabolism to withstand stresses such as low pH, reactive nitrogen species(RNS), reactive oxygen species(ROS) and several other anti-microbial effectors.Pathogen virulence determinants play significant roles in the buffering antimicrobial substances and determination the outcome of infection.Rv0341 open reading frame without defined function, that was shown to be upregulated in macrophages isolated from transition zone of human tuberculosis granuloma, upon in vitro infection of macrophages and under pressure of antibiotics targeting cell wall. Rv0341 has been proposed to be encode efflux pump and osmoprotective protein that can mediated antimicrobial resistance. To explore the function of M. tuberculosis Rv0341 in antimicrobial resistance and virulence of M. tuberculosis, Rv0341 was heterologously expressed in M. smegmatis.We found that overexpression of Rv0341 altered the bacterial colonial morphology, reduced sliding motility and biofilm formation. Rv0341 reduces susceptibility of M. smegmatis to glycopeptide antibiotics and in vitro stressors including low pH, SDS, H2O2 and diamide in comparison to M. smegmatis harboring vector only. The Ethidium bromide uptake was used for identification cell wall permeability in recombinant strains, we found that cell wall permeability in Ms_Rv0341 was significantly reduced compared to Ms_Vec strain. In addition, Rv0341 changes the genes expression patterns of M. smegmatis by upregulation genes involved in mycobacterial stress response and virulence. The growth of Rv0341 expressing strain was improved in the macrophages compared to the Ms_Vec strain. Colonization of macrophages by M. smegmatis expressing Rv0341 was associated significantly greater amount of TNF-alpha and IL-1beta compared with controls. The data firstly showed that Rv0341 plays a role in Mycobacterium physiology and pathogenesis via changing the cell wall permeability.