Investigating the composition and recruitment of the mycobacterial ImuA′–ImuB–DnaE2 mutasome
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 polym...
| Main Authors: | , , , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2023-08-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/75628 |
| _version_ | 1797745062880215040 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Sophia Gessner |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-12T15:18:08Z |
| format | Article |
| id | doaj.art-9340d5204dad4cac96d701c78d904d63 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-03-12T15:18:08Z |
| publishDate | 2023-08-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-9340d5204dad4cac96d701c78d904d632023-08-11T09:58:34ZengeLife Sciences Publications LtdeLife2050-084X2023-08-011210.7554/eLife.75628Investigating the composition and recruitment of the mycobacterial ImuA′–ImuB–DnaE2 mutasomeSophia Gessner0https://orcid.org/0000-0003-0824-0079Zela Alexandria-Mae Martin1Michael A Reiche2Joana A Santos3Ryan Dinkele4Atondaho Ramudzuli5Neeraj Dhar6https://orcid.org/0000-0002-5887-8137Timothy J de Wet7https://orcid.org/0000-0002-3978-5322Saber Anoosheh8Dirk M Lang9Jesse Aaron10Teng-Leong Chew11Jennifer Herrmann12Rolf Müller13https://orcid.org/0000-0002-1042-5665John D McKinney14https://orcid.org/0000-0002-0557-3479Roger Woodgate15https://orcid.org/0000-0001-5581-4616Valerie Mizrahi16https://orcid.org/0000-0003-4824-9115Česlovas Venclovas17Meindert H Lamers18https://orcid.org/0000-0002-4205-1338Digby F Warner19https://orcid.org/0000-0002-4146-0930SAMRC/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 AfricaSAMRC/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, SwitzerlandSAMRC/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 StatesDepartment of Cell and Chemical Biology, Leiden University Medical Center, Leiden, NetherlandsSAMRC/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 AfricaSAMRC/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 AfricaLaboratory of Microbiology and Microsystems, School of Life Sciences, Swiss Federal Institute of Technology in Lausanne (EPFL), Lausanne, SwitzerlandSAMRC/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 AfricaSAMRC/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 AfricaConfocal and Light Microscope Imaging Facility, Department of Human Biology, University of Cape Town, Cape Town, South AfricaAdvanced Imaging Center, Howard Hughes Medical Institute, Ashburn, United StatesAdvanced Imaging Center, Howard Hughes Medical Institute, Ashburn, United StatesHelmholtz Centre for Infection Research, Helmholtz Institute for Pharmaceutical Research Saarland, Saarbrücken, Germany; German Centre for Infection Research (DZIF), Partner Site Hannover-Braunschweig, Braunschweig, GermanyHelmholtz Centre for Infection Research, Helmholtz Institute for Pharmaceutical Research Saarland, Saarbrücken, Germany; German Centre for Infection Research (DZIF), Partner Site Hannover-Braunschweig, Braunschweig, GermanyLaboratory of Microbiology and Microsystems, School of Life Sciences, Swiss Federal Institute of Technology in Lausanne (EPFL), Lausanne, SwitzerlandLaboratory of Genomic Integrity, Eunice Kennedy Shriver National Institute of Child Health and Human Development, Bethesda, United StatesSAMRC/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 AfricaInstitute of Biotechnology, Vilnius University, Vilnius, LithuaniaDepartment of Cell and Chemical Biology, Leiden University Medical Center, Leiden, NetherlandsSAMRC/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 AfricaA 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.https://elifesciences.org/articles/75628Mycobacterium tuberculosisMycobacterium smegmatisinduced mutagenesisantibiotic resistanceanti-evolutionmutasome |
| spellingShingle | 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 Investigating the composition and recruitment of the mycobacterial ImuA′–ImuB–DnaE2 mutasome eLife Mycobacterium tuberculosis Mycobacterium smegmatis induced mutagenesis antibiotic resistance anti-evolution mutasome |
| title | Investigating the composition and recruitment of the mycobacterial ImuA′–ImuB–DnaE2 mutasome |
| title_full | Investigating the composition and recruitment of the mycobacterial ImuA′–ImuB–DnaE2 mutasome |
| title_fullStr | Investigating the composition and recruitment of the mycobacterial ImuA′–ImuB–DnaE2 mutasome |
| title_full_unstemmed | Investigating the composition and recruitment of the mycobacterial ImuA′–ImuB–DnaE2 mutasome |
| title_short | Investigating the composition and recruitment of the mycobacterial ImuA′–ImuB–DnaE2 mutasome |
| title_sort | investigating the composition and recruitment of the mycobacterial imua imub dnae2 mutasome |
| topic | Mycobacterium tuberculosis Mycobacterium smegmatis induced mutagenesis antibiotic resistance anti-evolution mutasome |
| url | https://elifesciences.org/articles/75628 |
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