Depletion of the DarG antitoxin in Mycobacterium tuberculosis triggers the DNA-damage response and leads to cell death

Abstract

Of the ~80 putative toxin-antitoxin (TA) modules encoded by the bacterial pathogen Mycobacterium tuberculosis (Mtb), three contain antitoxins essential for bacterial viability. One of these, Rv0060 (DNA ADP-ribosyl glycohydrolase, DarG_Mtb ), functions along with its cognate toxin Rv0059 (DNA ADP-ribosyl transferase, DarT_Mtb ), to mediate reversible DNA ADP-ribosylation (Jankevicius et al., 2016). We demonstrate that DarT_Mtb -DarG_Mtb form a functional TA pair and essentiality of darG_Mtb is dependent on the presence of darT_Mtb , but simultaneous deletion of both darT_Mtb -darG_Mtb does not alter viability of Mtb in vitro or in mice. The antitoxin, DarG_Mtb , forms a cytosolic complex with DNA-repair proteins that assembles independently of either DarT_Mtb or interaction with DNA. Depletion of DarG_Mtb alone is bactericidal, a phenotype that is rescued by expression of an orthologous antitoxin, DarG_Taq , from Thermus aquaticus. Partial depletion of DarG_Mtb triggers a DNA-damage response and sensitizes Mtb to drugs targeting DNA metabolism and respiration. Induction of the DNA-damage response is essential for Mtb to survive partial DarG_Mtb -depletion and leads to a hypermutable phenotype.

Keywords: Mycobacterium tuberculosis; DNA damage; toxin-antitoxin systems.