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. Author manuscript; available in PMC: 2017 Aug 23.
Published in final edited form as: Nat Microbiol. 2017 Jan 13;2:16253. doi: 10.1038/nmicrobiol.2016.253

Figure 2. Evidence for RodA as a candidate for the missing GTase.

Figure 2

a, Synthetic lethality of a rodA mutation in a ΔponA and Δ4 mutants. Growth on NA/MSM plates with or without 1 mM IPTG of strains YK2245 (Pspac-rodA), YK2246 (ΔponA::spc Pspac-rodA) and YK2242 (Δ4 Pspac-rodA) at 30°C.

b, Growth profiles of YK2245 (Pspac-rodA), YK2246 (ΔponA::spc Pspac-rodA) and YK2242 (Δ4 Pspac-rodA) strains in NB/MSM with or without 1 mM IPTG at 37°C.

c, Morphological effects of RodA depletion in ΔponA and Δ4 mutants. Phase-contrast and the corresponding cell membrane-stained images of typical fields of cells of YK2245 (Pspac-rodA), YK2246 (ΔponA::spc Pspac-rodA) and YK2242 (Δ4 Pspac-rodA) cultured in NB/MSM medium with or without 1 mM IPTG at 37°C. Time (h) of cultivation of cells after removal of IPTG is indicated. Scale bar represents 3 μm.

d, Rescue of the growth defect of a Δ4 mutant by overexpression of rodA, but not ftsW. Growth on PAB plates with or without 1 mM IPTG, or 1% xylose, of strains YK2256 (Δ4 pLOSS-Pspac-ponA amyE::Pxyl-rodA) and YK2257 (Δ4 pLOSS-Pspac-ponA amyE::Pxyl-ftsW) at 30°C.

e, Rescue of the synthetic lethality of Δ4 and ΔsigM strains by the overexpression of rodA. Growth on NA plates with or without 1 mM IPTG, or 1% xylose of strain YK2259 (Δ4 pLOSS-Pspac-ponA amyE::Pxyl-rodA ΔsigM::kan) at 30°C.

f, Restoration of resistance to MOE in a sigM mutant overproducing RodA. Wild type (168CA), AG494 (ΔsigM) or YK2268 (ΔsigM amyE::Pxyl-rodA) were grown in the presence (bottom) or absence (top) of xylose, and treated with MOE as indicated.

All the results in the figure are representative of at least two independent experiments.