Mycobacterium tuberculosis WhiB4 regulates oxidative stress response to modulate survival and dissemination in vivo

Manbeena Chawla, Pankti Parikh, Alka Saxena, Mohamedhusen Munshi, Mansi Mehta, Deborah Mai, Anup K. Srivastava, K. V. Narasimhulu, Kevin Redding, Nimi Vashi, Dhiraj Kumar, Adrie J C Steyn, Amit Singh

    Research output: Contribution to journalArticle

    41 Citations (Scopus)

    Abstract

    Host-generated oxidative stress is considered one of the main mechanisms constraining Mycobacterium tuberculosis (Mtb) growth. The redox-sensing mechanisms in Mtb are not completely understood. Here we show that WhiB4 responds to oxygen (O2) and nitric oxide (NO) via its 4Fe-4S cluster and controls the oxidative stress response in Mtb. The WhiB4 mutant (MtbΔwhiB4) displayed an altered redox balance and a reduced membrane potential. Microarray analysis demonstrated that MtbΔwhiB4 overexpresses the antioxidant systems including alkyl hydroperoxidase (ahpC-ahpD) and rubredoxins (rubA-rubB). DNA binding assays showed that WhiB4 [4Fe-4S] cluster is dispensable for DNA binding. However, oxidation of the apo-WhiB4 Cys thiols induced disulphide-linked oligomerization, DNA binding and transcriptional repression, whereas reduction reversed the effect. Furthermore, WhiB4 binds DNA with a preference for GC-rich sequences. Expression analysis showed that oxidative stress repressed whiB4 and induced antioxidants in Mtb, while their hyper-induction was observed in MtbΔwhiB4. MtbΔwhiB4 showed increased resistance to oxidative stress in vitro and enhanced survival inside the macrophages. Lastly, MtbΔwhiB4 displayed hypervirulence in the lungs of guinea pigs, but showed a defect in dissemination to their spleen. These findings suggest that WhiB4 systematically calibrates the activation of oxidative stress response in Mtb to maintain redox balance, and to modulate virulence.

    Original languageEnglish (US)
    Pages (from-to)1148-1165
    Number of pages18
    JournalMolecular Microbiology
    Volume85
    Issue number6
    DOIs
    StatePublished - Sep 2012

    Fingerprint

    Mycobacterium tuberculosis
    Oxidative Stress
    Oxidation-Reduction
    DNA
    Rubredoxins
    Antioxidants
    GC Rich Sequence
    Microarray Analysis
    Sulfhydryl Compounds
    Disulfides
    Membrane Potentials
    Virulence
    Nitric Oxide
    Guinea Pigs
    Spleen
    Macrophages
    Oxygen
    Lung
    Growth

    ASJC Scopus subject areas

    • Molecular Biology
    • Microbiology

    Cite this

    Mycobacterium tuberculosis WhiB4 regulates oxidative stress response to modulate survival and dissemination in vivo. / Chawla, Manbeena; Parikh, Pankti; Saxena, Alka; Munshi, Mohamedhusen; Mehta, Mansi; Mai, Deborah; Srivastava, Anup K.; Narasimhulu, K. V.; Redding, Kevin; Vashi, Nimi; Kumar, Dhiraj; Steyn, Adrie J C; Singh, Amit.

    In: Molecular Microbiology, Vol. 85, No. 6, 09.2012, p. 1148-1165.

    Research output: Contribution to journalArticle

    Chawla, M, Parikh, P, Saxena, A, Munshi, M, Mehta, M, Mai, D, Srivastava, AK, Narasimhulu, KV, Redding, K, Vashi, N, Kumar, D, Steyn, AJC & Singh, A 2012, 'Mycobacterium tuberculosis WhiB4 regulates oxidative stress response to modulate survival and dissemination in vivo' Molecular Microbiology, vol. 85, no. 6, pp. 1148-1165. https://doi.org/10.1111/j.1365-2958.2012.08165.x
    Chawla, Manbeena ; Parikh, Pankti ; Saxena, Alka ; Munshi, Mohamedhusen ; Mehta, Mansi ; Mai, Deborah ; Srivastava, Anup K. ; Narasimhulu, K. V. ; Redding, Kevin ; Vashi, Nimi ; Kumar, Dhiraj ; Steyn, Adrie J C ; Singh, Amit. / Mycobacterium tuberculosis WhiB4 regulates oxidative stress response to modulate survival and dissemination in vivo. In: Molecular Microbiology. 2012 ; Vol. 85, No. 6. pp. 1148-1165.
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    AU - Mehta, Mansi

    AU - Mai, Deborah

    AU - Srivastava, Anup K.

    AU - Narasimhulu, K. V.

    AU - Redding, Kevin

    AU - Vashi, Nimi

    AU - Kumar, Dhiraj

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