The identification Mycobacterium tuberculosis genes that modulate long term survival in the presence of rifampicin and streptomycin

Abstract In 2023, Mycobacterium tuberculosis (Mtb) caused 10.6 million new tuberculosis cases and 1.3 million deaths. The WHO proscribed treatment is not always successful, even when strains were sensitive to the antibiotics.as clinical Mtb populations contain phenotypically tolerant subpopulations,...

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Main Authors: Johana E. Hernández Toloza, Ye Xu, Tom A. Mendum, Bianca Sica Siedler, Rosalyn Casey, Huihai Wu, Kerstin Williams, Suzanne Hingley-Wilson, Johnjoe McFadden
Format: Article
Language:English
Published: Nature Portfolio 2025-07-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-04038-9
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author Johana E. Hernández Toloza
Ye Xu
Tom A. Mendum
Bianca Sica Siedler
Rosalyn Casey
Huihai Wu
Kerstin Williams
Suzanne Hingley-Wilson
Johnjoe McFadden
author_facet Johana E. Hernández Toloza
Ye Xu
Tom A. Mendum
Bianca Sica Siedler
Rosalyn Casey
Huihai Wu
Kerstin Williams
Suzanne Hingley-Wilson
Johnjoe McFadden
author_sort Johana E. Hernández Toloza
collection DOAJ
description Abstract In 2023, Mycobacterium tuberculosis (Mtb) caused 10.6 million new tuberculosis cases and 1.3 million deaths. The WHO proscribed treatment is not always successful, even when strains were sensitive to the antibiotics.as clinical Mtb populations contain phenotypically tolerant subpopulations, termed persisters. Here a Mtb transposon library was challenged with rifampicin (RIF) and streptomycin (STM) under conditions designed to identify genes that modulate persister frequency. Mutants with reduced survival in RIF were predominantly in genes associated with membrane integrity e.g. arabinogalactan assembly genes cpsA/lytR/Psr, whilst for STM, reduced survival was associated with toxin/antitoxin genes. Some mutations enhanced survival. For RIF these included the methyl citrate cycle genes prpC, prpD and prpR, and the trkA-C K+ uptake system genes ceoB and Rv2690, and for STM, the resistance associated gene, gidB, and anion-transport genes Rv3679c and Rv3680c. Few genes overlapped the RIF and STM selections, demonstrating that survival mechanisms were antibiotic-specific. Directed deletions of ΔprpD and ΔfadE5 confirmed their predicted enhanced and reduced RIF fitness respectively. The study identified genes that modulate not only persister frequency but also resistance and tolerance, and demonstrates that the mechanisms that produce these phenotypes are diverse and antibiotic-specific.
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spelling doaj-art-b7b0ec1a43ca44aeb06edd13aaee4cd42025-08-20T04:01:36ZengNature PortfolioScientific Reports2045-23222025-07-0115111510.1038/s41598-025-04038-9The identification Mycobacterium tuberculosis genes that modulate long term survival in the presence of rifampicin and streptomycinJohana E. Hernández Toloza0Ye Xu1Tom A. Mendum2Bianca Sica Siedler3Rosalyn Casey4Huihai Wu5Kerstin Williams6Suzanne Hingley-Wilson7Johnjoe McFadden8Department of Microbial Sciences, School of Biosciences, Faculty of Health and Medical Sciences, University of SurreyDepartment of Microbial Sciences, School of Biosciences, Faculty of Health and Medical Sciences, University of SurreyDepartment of Microbial Sciences, School of Biosciences, Faculty of Health and Medical Sciences, University of SurreyDepartment of Microbial Sciences, School of Biosciences, Faculty of Health and Medical Sciences, University of SurreyDepartment of Microbial Sciences, School of Biosciences, Faculty of Health and Medical Sciences, University of SurreyBioinformatics Facility, Faculty of Health and Medical Sciences, University of SurreyDepartment of Microbial Sciences, School of Biosciences, Faculty of Health and Medical Sciences, University of SurreyDepartment of Microbial Sciences, School of Biosciences, Faculty of Health and Medical Sciences, University of SurreyDepartment of Microbial Sciences, School of Biosciences, Faculty of Health and Medical Sciences, University of SurreyAbstract In 2023, Mycobacterium tuberculosis (Mtb) caused 10.6 million new tuberculosis cases and 1.3 million deaths. The WHO proscribed treatment is not always successful, even when strains were sensitive to the antibiotics.as clinical Mtb populations contain phenotypically tolerant subpopulations, termed persisters. Here a Mtb transposon library was challenged with rifampicin (RIF) and streptomycin (STM) under conditions designed to identify genes that modulate persister frequency. Mutants with reduced survival in RIF were predominantly in genes associated with membrane integrity e.g. arabinogalactan assembly genes cpsA/lytR/Psr, whilst for STM, reduced survival was associated with toxin/antitoxin genes. Some mutations enhanced survival. For RIF these included the methyl citrate cycle genes prpC, prpD and prpR, and the trkA-C K+ uptake system genes ceoB and Rv2690, and for STM, the resistance associated gene, gidB, and anion-transport genes Rv3679c and Rv3680c. Few genes overlapped the RIF and STM selections, demonstrating that survival mechanisms were antibiotic-specific. Directed deletions of ΔprpD and ΔfadE5 confirmed their predicted enhanced and reduced RIF fitness respectively. The study identified genes that modulate not only persister frequency but also resistance and tolerance, and demonstrates that the mechanisms that produce these phenotypes are diverse and antibiotic-specific.https://doi.org/10.1038/s41598-025-04038-9Mycobacterium tuberculosisTransposon libraryTolerancePersisterCell wallMethylcitrate cycle
spellingShingle Johana E. Hernández Toloza
Ye Xu
Tom A. Mendum
Bianca Sica Siedler
Rosalyn Casey
Huihai Wu
Kerstin Williams
Suzanne Hingley-Wilson
Johnjoe McFadden
The identification Mycobacterium tuberculosis genes that modulate long term survival in the presence of rifampicin and streptomycin
Scientific Reports
Mycobacterium tuberculosis
Transposon library
Tolerance
Persister
Cell wall
Methylcitrate cycle
title The identification Mycobacterium tuberculosis genes that modulate long term survival in the presence of rifampicin and streptomycin
title_full The identification Mycobacterium tuberculosis genes that modulate long term survival in the presence of rifampicin and streptomycin
title_fullStr The identification Mycobacterium tuberculosis genes that modulate long term survival in the presence of rifampicin and streptomycin
title_full_unstemmed The identification Mycobacterium tuberculosis genes that modulate long term survival in the presence of rifampicin and streptomycin
title_short The identification Mycobacterium tuberculosis genes that modulate long term survival in the presence of rifampicin and streptomycin
title_sort identification mycobacterium tuberculosis genes that modulate long term survival in the presence of rifampicin and streptomycin
topic Mycobacterium tuberculosis
Transposon library
Tolerance
Persister
Cell wall
Methylcitrate cycle
url https://doi.org/10.1038/s41598-025-04038-9
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