Zn Intoxication of Mycobacterium marinum during Dictyostelium discoideum Infection Is Counteracted by Induction of the Pathogen Zn Exporter CtpC.

Macrophages use diverse strategies to restrict intracellular pathogens, including either depriving the bacteria of (micro)nutrients such as transition metals or intoxicating them via metal accumulation. Little is known about the chemical warfare between , a close relative of (Mtb), and its hosts. We use the professional phagocyte to investigate the role of Zn during infection. We show that senses toxic levels of Zn and responds by upregulating one of its isoforms of the Zn efflux transporter CtpC. Deletion of (MMAR_1271) leads to growth inhibition in broth supplemented with Zn as well as reduced intracellular growth. Both phenotypes were fully rescued by constitutive ectopic expression of the Mtb CtpC orthologue demonstrating that MMAR_1271 is the functional CtpC Zn efflux transporter in Infection leads to the accumulation of Zn inside the -containing vacuole (MCV), achieved by the induction and recruitment of the Zn efflux pumps ZntA and ZntB. In cells lacking ZntA, there is further attenuation of growth, presumably due to a compensatory efflux of Zn into the MCV, carried out by ZntB, the main Zn transporter in endosomes and phagosomes. Counterintuitively, bacterial growth is also impaired in KO cells, in which MCVs appear to accumulate less Zn than in wild-type cells, suggesting restriction by other Zn-mediated mechanisms. Absence of CtpC further epistatically attenuates the intracellular proliferation of in and KO cells, confirming that mycobacteria face noxious levels of Zn Microelements are essential for the function of the innate immune system. A deficiency in zinc or copper results in an increased susceptibility to bacterial infections. Zn serves as an important catalytic and structural cofactor for a variety of enzymes including transcription factors and enzymes involved in cell signaling. But Zn is toxic at high concentrations and represents a cell-autonomous immunity strategy that ensures killing of intracellular bacteria in a process called zinc poisoning. The cytosolic and lumenal Zn concentrations result from the balance of import into the cytosol via ZIP influx transporters and efflux via ZnT transporters. Here, we show that Zn poisoning is involved in restricting infections. Our study extends observations during infection and explores for the first time how the interplay of ZnT transporters affects mycobacterial infection by impacting Zn homeostasis.