Fig. 5: Ribosome biogenesis is more vulnerable to inhibition in an INHR-katG mutant. | Nature Communications

Fig. 5: Ribosome biogenesis is more vulnerable to inhibition in an INHR-katG mutant.

From: Whole genome CRISPRi screening identifies druggable vulnerabilities in an isoniazid resistant strain of Mycobacterium tuberculosis

Fig. 5

a Pathway analysis of more vulnerable genes in INHR-katG within the protein synthesis class. Subclasses are described using classifications from the curated PATRIC database. The bubble plot represents data from day 14. The x-axis quantifies the proportion of genes called “more vulnerable” over the total number of M. tuberculosis genes per functional subclass of the protein synthesis pathway, and the y-axis shows the name of each functional subclass and the total number of genes in each subclass. Within each functional subclass, the dot size indicates the average ratio of gRNAs targeting each gene that is more depleted in INHR-katG. The dot colour denotes the ATc concentration from which the amplicon sequencing was performed. Ribo-bio: Ribosome biogenesis. Aa-tRNA: Aminoacyl-tRNA-synthetases. b Growth of M. tuberculosis DS-parent and INHR-katG expressing gRNAs targeting rplC in ATc dose response assays (mean ± extrema of two biological replicates, n=3 independent experiments). The (gx) after each gRNA denotes the specific gRNA targeting each gene. c, d M. tuberculosis DS-parent and INHR-katG expressing a gRNA targeting rplC and assessed (c) in continuous log phase growth in 7H9 media with ATc-300, (d) for viability in ATc dose response assays and (e) for intracellular survival within THP-1 macrophage cells (mean ± SD of three biological replicates, n = 2 independent experiments). Data presentation is consistent with Fig. 4 and experiments were performed as described in material and methods. f–h Susceptibility of M. tuberculosis DS-parent and INHR-katG to increasing concentrations of LZD as assessed (f) in 96 well plates, (g) in time kill assays and (h) against intracellular M. tuberculosis within THP-1 infected macrophages. For h, the MIC of LZD was 1 µM. For (f) Inoc denotes the starting CFU/ml and no-cpd denotes the detected CFU/ml in the absence of compound (mean ± SD of three biological replicates, n = 2 independent experiments). Dashed line represents the lower limit of detection. i M. tuberculosis DS-parent grown with INH (9 µM) with or without increasing concentrations of LZD (mean ± SD of three biological replicates, n = 2). Source data are provided as a Source Data file.

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