Naturally occurring dinactin targets cpsA protein and kills Mycobacterium tuberculosis by disrupting the proton motive force

Tuberculosis, especially drug-resistant tuberculosis remains a global threat, and new drugs are desperately needed to combat the spread of multidrug-resistant Mycobacterium tuberculosis. Here we describe a natural macrotetrolide dinactin with anti-tuberculosis activity against susceptive and non-replicating Mycobacterium tuberculosis. Dinactin can also synergistically enhance the anti-tuberculosis effect of rifampicin and isoniazid against drug-resistant strains.Furthermore, dinactin exhibited excellently antituberculosis effect in macrophage and Galleria mellonella models. Since the ionophore properties of dinactin, it not only enhanced cations transport and altered membrane permeability but also caused the dissipation of proton motive force and metabolic perturbations. Finally, through the selection of spontaneous resistant mutants and whole genome sequencing, non-synonymous single nucleotide polymorphisms were successfully identified in the cpsA gene of the LytR-Cps2A-Psr family. The dinactin-resistant mutants exhibited decreased in vitro drug sensitivity to dinactin without cross-resistance to first-line antituberculosis drugs. Genetic studies and molecular biology assays have subsequently confirmed cpsA as one of the potential targets for dinactin's anti-tuberculosis activity. Collectively, these data indicate that dinactin could be a promising candidate for treating tuberculosis.