eLife (Aug 2023)

Investigating the composition and recruitment of the mycobacterial ImuA′–ImuB–DnaE2 mutasome

  • Sophia Gessner,
  • Zela Alexandria-Mae Martin,
  • Michael A Reiche,
  • Joana A Santos,
  • Ryan Dinkele,
  • Atondaho Ramudzuli,
  • Neeraj Dhar,
  • Timothy J de Wet,
  • Saber Anoosheh,
  • Dirk M Lang,
  • Jesse Aaron,
  • Teng-Leong Chew,
  • Jennifer Herrmann,
  • Rolf Müller,
  • John D McKinney,
  • Roger Woodgate,
  • Valerie Mizrahi,
  • Česlovas Venclovas,
  • Meindert H Lamers,
  • Digby F Warner

DOI
https://doi.org/10.7554/eLife.75628
Journal volume & issue
Vol. 12

Abstract

Read online

A DNA damage-inducible mutagenic gene cassette has been implicated in the emergence of drug resistance in Mycobacterium tuberculosis during anti-tuberculosis (TB) chemotherapy. However, the molecular composition and operation of the encoded ‘mycobacterial mutasome’ – minimally comprising DnaE2 polymerase and ImuA′ and ImuB accessory proteins – remain elusive. Following exposure of mycobacteria to DNA damaging agents, we observe that DnaE2 and ImuB co-localize with the DNA polymerase III β subunit (β clamp) in distinct intracellular foci. Notably, genetic inactivation of the mutasome in an imuBAAAAGG mutant containing a disrupted β clamp-binding motif abolishes ImuB–β clamp focus formation, a phenotype recapitulated pharmacologically by treating bacilli with griselimycin and in biochemical assays in which this β clamp-binding antibiotic collapses pre-formed ImuB–β clamp complexes. These observations establish the essentiality of the ImuB–β clamp interaction for mutagenic DNA repair in mycobacteria, identifying the mutasome as target for adjunctive therapeutics designed to protect anti-TB drugs against emerging resistance.

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