Substrate recognition and cryo-EM structure of the ribosome-bound TAC toxin of Mycobacterium tuberculosis

Toxins of toxin-antitoxin systems use diverse mechanisms to control bacterial growth. Here, we focus on the deleterious toxin of the atypical tripartite toxin-antitoxin-chaperone (TAC) system of Mycobacterium tuberculosis, whose inhibition requires the concerted action of the antitoxin and its dedicated SecB-like chaperone. We show that the TAC toxin is a bona fide ribonuclease and identify exact cleavage sites in mRNA targets on a transcriptome-wide scale in vivo. mRNA cleavage by the toxin occurs after the second nucleotide of the ribosomal A-site codon during translation, with a strong preference for CCA codons in vivo. Finally, we report the cryo-EM structure of the ribosome-bound TAC toxin in the presence of native M. tuberculosis cspA mRNA, revealing the specific mechanism by which the TAC toxin interacts with the ribosome and the tRNA in the P-site to cleave its mRNA target. Toxin-antitoxin systems are widespread in bacteria. Here the authors present structures of M. tuberculosis HigB(TAC) alone and bound to the ribosome in the presence of native cspA mRNA, shedding light on its mechanism of translation inhibition.

Automated summary

This study reveals the mechanism of action of the toxin in the Mycobacterium tuberculosis TAC system. The researchers discovered that the TAC toxin is a genuine ribonuclease and identified the cleavage sites on mRNA targets on a transcriptome-wide scale. Additionally, they determined the specific mechanism by which the TAC toxin interacts with the ribosome and tRNA through cryo-electron microscopy.