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. Author manuscript; available in PMC: 2023 Sep 27.
Published in final edited form as: Mol Microbiol. 2023 Jul 14;120(3):351–383. doi: 10.1111/mmi.15122

Figure 6. Overproduction or absence of MurZ(Spn), but not MurA(Spn), alters growth, morphology, and sensitivity to fosfomycin or penicillin.

Figure 6.

(A and B) Parent D39 Δcps rpsL1 strain (IU1824), constructed murZ(D280Y) (IU13438), ΔmurZ (IU13536), and merodiploid murZ+//PZn-murZ+ (IU13393) strains were grown overnight in BHI broth with no additional (Zn2+/(1/10)Mn2+) and diluted to OD620 ≈0.003 in the morning in BHI broth with or without (Zn2+/(1/10)Mn2+) at the concentrations indicated. (A) Representative growth curves and averaged DT and yields. ** p< 0.01; *** p < 0.001 compared to WT strain by one-way ANOVA analysis (GraphPad Prism, Dunnett’s test). (B) Representative phase-contrast images taken between 3.5 to 4 h of growth for all strains and conditions, except for IU13393 with 0.4 mM (Zn2+(1/10)Mn2+), which was taken at 5 h of growth. (C and D) Parent D39 Δcps rpsL1 strain (IU1824), ΔmurA (IU13538), and merodiploid murA+//PZn-murA+ (IU13395) strains were grown similarly to the murZ strains described above. The DTs and growth yields of all strains and conditions were not statistically different from the values obtained for the WT strain. (D) Representative phase-contrast images taken at 3 h of growth for all strains and conditions. All micrographs in (B) and (D) are at the same magnification (scale bar = 1 μm). Box-and-whisker plots of cell dimensions of murZ(D280Y) and strains overexpressing murZ or murA are in Fig. S10. (E) Disc diffusion assays were performed as described in Experimental procedures for strains: WT parent (IU1824), ΔmurZ (IU13536), ΔmurA (IU13538), murZ(D280Y) (IU13438), murZ(C116S) (IU15939), murZ(D280Y) ΔmurA (IU17748), ΔkhpA (IU9036), ΔkhpA ΔmurZ (IU13542), ΔkhpA ΔmurA (IU13546), ΔclpP (IU12462), murZ+//PZn-murZ+ (no Zn) (IU13393), and murZ+//PZn-murZ+ in 0.2 mM (Zn2+/(1/10)Mn2+). Mean diameters of zones of inhibition ± SEM are graphed from at least two independent biological replicates. Means and numbers of replicates (n) are shown at the tops and bottoms of bars, respectively. P values were obtained by the Welch t-test (GraphPad Prism). *, **, and *** denote p<0.05, p<0.01, p<0.001, respectively.