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Mechanisms of Resistance

Resuscitation-Promoting Factors Are Required for β-Lactam Tolerance and the Permeability Barrier in Mycobacterium tuberculosis

Carl N. Wivagg, Deborah T. Hung
Carl N. Wivagg
aDepartment of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts, USA
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Deborah T. Hung
aDepartment of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts, USA
bInfectious Disease Initiative, Broad Institute, Cambridge, Massachusetts, USA
cDepartment of Molecular Biology and Center for Computational and Integrative Biology, Massachusetts General Hospital, Boston, Massachusetts, USA
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DOI: 10.1128/AAC.06027-11
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    Fig 1

    RPFWT and RPFnull were grown in 3 μM cefotaxime, and surviving bacteria were enumerated at the indicated time points by plating for CFU on 7H10 agar. The result shown is representative of the experiment performed in biological triplicate.

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    Fig 2

    Kinetics of cefamandole hydrolysis. Absorption at 285 nm of H37Rv (●) or H37RvΔrpfACDEB (■). Samples were mechanically lysed (A) or resuspended in phosphate-buffered saline intact (B) and exposed to 20 mM cefamandole nafate. Subsequently, absorption measurements were taken at the indicated times. The illustrated values are averages of two technical replicates for each strain. Parameters vlysed and vintact are the slope and error calculated from the aggregated data for each strain.

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    Additional hits from RPFnull screen

    Table 1
    • a ChemBank is a public cheminformatics resource available at http://chembank.broad.harvard.edu/ (7).

  • Table 2

    IC90s of β-lactams against H37Rv, RPFnull, and related strains

    β-LactamStrainaIC90 (μM)b
    CefotaximeH37Rv (RPFWT)26
    H37RvΔrpfA13–26
    H37RvΔrpfB13–26
    H37RvΔrpfC26
    H37RvΔrpfD26
    H37RvΔrpfE26–52
    H37RvΔrpfAB26
    H37RvΔrpfACBED::pHrpfCDE6.5–26
    H37RvΔrpfACBE6.5
    H37RvΔrpfACBED (RPFnull)1.6–3.3
    H37RvΔrpfACDEBc0.78
    CefamandoleH37Rv (RPFWT)390–780
    H37RvΔrpfACBED (RPFnull)98
        With potassium clavulanate (1 μM)H37Rv (RPFWT)24
    H37RvΔrpfACBED (RPFnull)1.5
    CeftriaxoneH37Rv (RPFWT)3.2–6.3
    H37RvΔrpfACBED (RPFnull)1.6
    • ↵a From reference 5.

    • ↵b Differences in IC90s are representative of at least two independent experiments containing at least four duplicates each; for cefamandole, amounts of clavulanate were varied across duplicates.

    • ↵c RPFs were deleted from this strain in a different order than that for RPFnull.

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Resuscitation-Promoting Factors Are Required for β-Lactam Tolerance and the Permeability Barrier in Mycobacterium tuberculosis
Carl N. Wivagg, Deborah T. Hung
Antimicrobial Agents and Chemotherapy Feb 2012, 56 (3) 1591-1594; DOI: 10.1128/AAC.06027-11

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Resuscitation-Promoting Factors Are Required for β-Lactam Tolerance and the Permeability Barrier in Mycobacterium tuberculosis
Carl N. Wivagg, Deborah T. Hung
Antimicrobial Agents and Chemotherapy Feb 2012, 56 (3) 1591-1594; DOI: 10.1128/AAC.06027-11
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