Cystathionine ?-Lyase is a component of cystine-mediated oxidative defense in lactobacillus reuteri br11

R LO, M TURNER, D BARRY, R SREEKUMAR, T WALSH, Philip Giffard

    Research output: Contribution to journalArticleResearchpeer-review

    Abstract

    Lactobacillus reuteri BR11 possesses a novel mechanism of oxidative defense involving an abundant cystine ABC transporter encoded by the cyuABC gene cluster. Large amounts of thiols, including H2S, are secreted upon cystine uptake by the CyuC transporter. A cystathionine ?-lyase (cgl) gene is cotranscribed with the cyu genes in several L. reuteri strains and was hypothesized to participate in cystine-mediated oxidative defense by producing reducing equivalents. This hypothesis was tested with L. reuteri BR11 by constructing a cgl mutant (PNG901) and comparing it to a similarly constructed cyuC mutant (PNG902). Although Cgl was required for H2S production from cystine, it was not crucial for oxidative defense in de Mann-Rogosa-Sharpe medium, in contrast to CyuC, whose inactivation resulted in lag-phase arrest in aerated cultures. The importance of Cgl in oxidative defense was seen only in the presence of hemin, which poses severe oxidative stress. The growth defects in aerated cultures of both mutants were alleviated by supplementation with cysteine (and cystine in the cgl mutant) but not methionine, with the cyuC mutant showing a much higher concentration requirement. We conclude that L. reuteri BR11 requires a high concentration of exogenous cysteine/cystine to grow optimally under aerobic conditions. This requirement is fulfilled by the abundant CyuC transporter, which has probably arisen due to the broad substrate specificity of Cgl, resulting in a futile pathway which degrades cystine taken up by the CyuC transporter to H2S. Cgl plays a secondary role in oxidative defense by its well-documented function of cysteine biosynthesis. Copyright � 2009, American Society for Microbiology. All Rights Reserved.
    Original languageEnglish
    Pages (from-to)1827-1837
    Number of pages11
    JournalJournal of Bacteriology
    Volume191
    Issue number6
    Publication statusPublished - 2009

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    Lactobacillus reuteri
    Cystathionine
    Lyases
    Cystine
    Cysteine
    Hemin
    ATP-Binding Cassette Transporters
    Multigene Family
    Substrate Specificity
    Microbiology
    Sulfhydryl Compounds
    Methionine
    Contrast Media
    Genes
    Oxidative Stress

    Cite this

    LO, R ; TURNER, M ; BARRY, D ; SREEKUMAR, R ; WALSH, T ; Giffard, Philip. / Cystathionine ?-Lyase is a component of cystine-mediated oxidative defense in lactobacillus reuteri br11. In: Journal of Bacteriology. 2009 ; Vol. 191, No. 6. pp. 1827-1837.
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    abstract = "Lactobacillus reuteri BR11 possesses a novel mechanism of oxidative defense involving an abundant cystine ABC transporter encoded by the cyuABC gene cluster. Large amounts of thiols, including H2S, are secreted upon cystine uptake by the CyuC transporter. A cystathionine ?-lyase (cgl) gene is cotranscribed with the cyu genes in several L. reuteri strains and was hypothesized to participate in cystine-mediated oxidative defense by producing reducing equivalents. This hypothesis was tested with L. reuteri BR11 by constructing a cgl mutant (PNG901) and comparing it to a similarly constructed cyuC mutant (PNG902). Although Cgl was required for H2S production from cystine, it was not crucial for oxidative defense in de Mann-Rogosa-Sharpe medium, in contrast to CyuC, whose inactivation resulted in lag-phase arrest in aerated cultures. The importance of Cgl in oxidative defense was seen only in the presence of hemin, which poses severe oxidative stress. The growth defects in aerated cultures of both mutants were alleviated by supplementation with cysteine (and cystine in the cgl mutant) but not methionine, with the cyuC mutant showing a much higher concentration requirement. We conclude that L. reuteri BR11 requires a high concentration of exogenous cysteine/cystine to grow optimally under aerobic conditions. This requirement is fulfilled by the abundant CyuC transporter, which has probably arisen due to the broad substrate specificity of Cgl, resulting in a futile pathway which degrades cystine taken up by the CyuC transporter to H2S. Cgl plays a secondary role in oxidative defense by its well-documented function of cysteine biosynthesis. Copyright � 2009, American Society for Microbiology. All Rights Reserved.",
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    author = "R LO and M TURNER and D BARRY and R SREEKUMAR and T WALSH and Philip Giffard",
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    LO, R, TURNER, M, BARRY, D, SREEKUMAR, R, WALSH, T & Giffard, P 2009, 'Cystathionine ?-Lyase is a component of cystine-mediated oxidative defense in lactobacillus reuteri br11', Journal of Bacteriology, vol. 191, no. 6, pp. 1827-1837.

    Cystathionine ?-Lyase is a component of cystine-mediated oxidative defense in lactobacillus reuteri br11. / LO, R; TURNER, M; BARRY, D; SREEKUMAR, R; WALSH, T; Giffard, Philip.

    In: Journal of Bacteriology, Vol. 191, No. 6, 2009, p. 1827-1837.

    Research output: Contribution to journalArticleResearchpeer-review

    TY - JOUR

    T1 - Cystathionine ?-Lyase is a component of cystine-mediated oxidative defense in lactobacillus reuteri br11

    AU - LO, R

    AU - TURNER, M

    AU - BARRY, D

    AU - SREEKUMAR, R

    AU - WALSH, T

    AU - Giffard, Philip

    PY - 2009

    Y1 - 2009

    N2 - Lactobacillus reuteri BR11 possesses a novel mechanism of oxidative defense involving an abundant cystine ABC transporter encoded by the cyuABC gene cluster. Large amounts of thiols, including H2S, are secreted upon cystine uptake by the CyuC transporter. A cystathionine ?-lyase (cgl) gene is cotranscribed with the cyu genes in several L. reuteri strains and was hypothesized to participate in cystine-mediated oxidative defense by producing reducing equivalents. This hypothesis was tested with L. reuteri BR11 by constructing a cgl mutant (PNG901) and comparing it to a similarly constructed cyuC mutant (PNG902). Although Cgl was required for H2S production from cystine, it was not crucial for oxidative defense in de Mann-Rogosa-Sharpe medium, in contrast to CyuC, whose inactivation resulted in lag-phase arrest in aerated cultures. The importance of Cgl in oxidative defense was seen only in the presence of hemin, which poses severe oxidative stress. The growth defects in aerated cultures of both mutants were alleviated by supplementation with cysteine (and cystine in the cgl mutant) but not methionine, with the cyuC mutant showing a much higher concentration requirement. We conclude that L. reuteri BR11 requires a high concentration of exogenous cysteine/cystine to grow optimally under aerobic conditions. This requirement is fulfilled by the abundant CyuC transporter, which has probably arisen due to the broad substrate specificity of Cgl, resulting in a futile pathway which degrades cystine taken up by the CyuC transporter to H2S. Cgl plays a secondary role in oxidative defense by its well-documented function of cysteine biosynthesis. Copyright � 2009, American Society for Microbiology. All Rights Reserved.

    AB - Lactobacillus reuteri BR11 possesses a novel mechanism of oxidative defense involving an abundant cystine ABC transporter encoded by the cyuABC gene cluster. Large amounts of thiols, including H2S, are secreted upon cystine uptake by the CyuC transporter. A cystathionine ?-lyase (cgl) gene is cotranscribed with the cyu genes in several L. reuteri strains and was hypothesized to participate in cystine-mediated oxidative defense by producing reducing equivalents. This hypothesis was tested with L. reuteri BR11 by constructing a cgl mutant (PNG901) and comparing it to a similarly constructed cyuC mutant (PNG902). Although Cgl was required for H2S production from cystine, it was not crucial for oxidative defense in de Mann-Rogosa-Sharpe medium, in contrast to CyuC, whose inactivation resulted in lag-phase arrest in aerated cultures. The importance of Cgl in oxidative defense was seen only in the presence of hemin, which poses severe oxidative stress. The growth defects in aerated cultures of both mutants were alleviated by supplementation with cysteine (and cystine in the cgl mutant) but not methionine, with the cyuC mutant showing a much higher concentration requirement. We conclude that L. reuteri BR11 requires a high concentration of exogenous cysteine/cystine to grow optimally under aerobic conditions. This requirement is fulfilled by the abundant CyuC transporter, which has probably arisen due to the broad substrate specificity of Cgl, resulting in a futile pathway which degrades cystine taken up by the CyuC transporter to H2S. Cgl plays a secondary role in oxidative defense by its well-documented function of cysteine biosynthesis. Copyright � 2009, American Society for Microbiology. All Rights Reserved.

    KW - ABC transporter

    KW - carrier protein

    KW - cystathionine gamma lyase

    KW - cysteine

    KW - cystine

    KW - hemin

    KW - hydrogen sulfide

    KW - methionine

    KW - protein CyuC

    KW - thiol derivative

    KW - unclassified drug

    KW - bacterial protein

    KW - amino acid synthesis

    KW - article

    KW - bacterial gene

    KW - bacterial growth

    KW - bacterium culture

    KW - bacterium mutant

    KW - cgl gene

    KW - controlled study

    KW - cyuC gene

    KW - enzyme specificity

    KW - gene cluster

    KW - gene inactivation

    KW - genetic transcription

    KW - Lactobacillus reuteri

    KW - nonhuman

    KW - nucleotide sequence

    KW - oxidative stress

    KW - priority journal

    KW - reduction

    KW - enzymology

    KW - genetics

    KW - metabolism

    KW - molecular genetics

    KW - oxidation reduction reaction

    KW - Bacterial Proteins

    KW - Cystathionine gamma-Lyase

    KW - Cysteine

    KW - Cystine

    KW - Membrane Transport Proteins

    KW - Molecular Sequence Data

    KW - Oxidation-Reduction

    KW - Oxidative Stress

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    M3 - Article

    VL - 191

    SP - 1827

    EP - 1837

    JO - Journal of Bacteriology

    JF - Journal of Bacteriology

    SN - 0021-9193

    IS - 6

    ER -