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
Affiliations
- Sophia Gessner
- ORCiD
- SAMRC/NHLS/UCT Molecular Mycobacteriology Research Unit, DSI/NRF Centre of Excellence for Biomedical TB Research, Department of Pathology, University of Cape Town, Cape Town, South Africa; Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Cape Town, South Africa
- Zela Alexandria-Mae Martin
- SAMRC/NHLS/UCT Molecular Mycobacteriology Research Unit, DSI/NRF Centre of Excellence for Biomedical TB Research, Department of Pathology, University of Cape Town, Cape Town, South Africa; Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Cape Town, South Africa; Laboratory of Microbiology and Microsystems, School of Life Sciences, Swiss Federal Institute of Technology in Lausanne (EPFL), Lausanne, Switzerland
- Michael A Reiche
- SAMRC/NHLS/UCT Molecular Mycobacteriology Research Unit, DSI/NRF Centre of Excellence for Biomedical TB Research, Department of Pathology, University of Cape Town, Cape Town, South Africa; Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Cape Town, South Africa; Advanced Imaging Center, Howard Hughes Medical Institute, Ashburn, United States
- Joana A Santos
- Department of Cell and Chemical Biology, Leiden University Medical Center, Leiden, Netherlands
- Ryan Dinkele
- SAMRC/NHLS/UCT Molecular Mycobacteriology Research Unit, DSI/NRF Centre of Excellence for Biomedical TB Research, Department of Pathology, University of Cape Town, Cape Town, South Africa; Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Cape Town, South Africa
- Atondaho Ramudzuli
- SAMRC/NHLS/UCT Molecular Mycobacteriology Research Unit, DSI/NRF Centre of Excellence for Biomedical TB Research, Department of Pathology, University of Cape Town, Cape Town, South Africa; Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Cape Town, South Africa
- Neeraj Dhar
- ORCiD
- Laboratory of Microbiology and Microsystems, School of Life Sciences, Swiss Federal Institute of Technology in Lausanne (EPFL), Lausanne, Switzerland
- Timothy J de Wet
- ORCiD
- SAMRC/NHLS/UCT Molecular Mycobacteriology Research Unit, DSI/NRF Centre of Excellence for Biomedical TB Research, Department of Pathology, University of Cape Town, Cape Town, South Africa; Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Cape Town, South Africa; Department of Integrative Biomedical Sciences, University of Cape Town, Cape Town, South Africa
- Saber Anoosheh
- SAMRC/NHLS/UCT Molecular Mycobacteriology Research Unit, DSI/NRF Centre of Excellence for Biomedical TB Research, Department of Pathology, University of Cape Town, Cape Town, South Africa; Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Cape Town, South Africa
- Dirk M Lang
- Confocal and Light Microscope Imaging Facility, Department of Human Biology, University of Cape Town, Cape Town, South Africa
- Jesse Aaron
- Advanced Imaging Center, Howard Hughes Medical Institute, Ashburn, United States
- Teng-Leong Chew
- Advanced Imaging Center, Howard Hughes Medical Institute, Ashburn, United States
- Jennifer Herrmann
- Helmholtz Centre for Infection Research, Helmholtz Institute for Pharmaceutical Research Saarland, Saarbrücken, Germany; German Centre for Infection Research (DZIF), Partner Site Hannover-Braunschweig, Braunschweig, Germany
- Rolf Müller
- ORCiD
- Helmholtz Centre for Infection Research, Helmholtz Institute for Pharmaceutical Research Saarland, Saarbrücken, Germany; German Centre for Infection Research (DZIF), Partner Site Hannover-Braunschweig, Braunschweig, Germany
- John D McKinney
- ORCiD
- Laboratory of Microbiology and Microsystems, School of Life Sciences, Swiss Federal Institute of Technology in Lausanne (EPFL), Lausanne, Switzerland
- Roger Woodgate
- ORCiD
- Laboratory of Genomic Integrity, Eunice Kennedy Shriver National Institute of Child Health and Human Development, Bethesda, United States
- Valerie Mizrahi
- ORCiD
- SAMRC/NHLS/UCT Molecular Mycobacteriology Research Unit, DSI/NRF Centre of Excellence for Biomedical TB Research, Department of Pathology, University of Cape Town, Cape Town, South Africa; Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Cape Town, South Africa; Wellcome Centre for Infectious Diseases Research in Africa, University of Cape Town, Cape Town, South Africa
- Česlovas Venclovas
- Institute of Biotechnology, Vilnius University, Vilnius, Lithuania
- Meindert H Lamers
- ORCiD
- Department of Cell and Chemical Biology, Leiden University Medical Center, Leiden, Netherlands
- Digby F Warner
- ORCiD
- SAMRC/NHLS/UCT Molecular Mycobacteriology Research Unit, DSI/NRF Centre of Excellence for Biomedical TB Research, Department of Pathology, University of Cape Town, Cape Town, South Africa; Institute of Infectious Disease and Molecular Medicine, University of Cape Town, Cape Town, South Africa; Wellcome Centre for Infectious Diseases Research in Africa, University of Cape Town, Cape Town, South Africa
- DOI
- https://doi.org/10.7554/eLife.75628
- Journal volume & issue
-
Vol. 12
Abstract
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.
Keywords
- Mycobacterium tuberculosis
- Mycobacterium smegmatis
- induced mutagenesis
- antibiotic resistance
- anti-evolution
- mutasome