Fig 5. Hypomorphic mutants in the PG precursor synthesis pathway rescue ΔdacA1 growth but not morphology.

(A) Indicated strains are chromosomal replacements of murA and murD with murA^P122S, murA^L35F and murD^D447E, respectively. Overnight cultures were plated on LB medium by spot- titering and incubated at 30°C overnight. (B) Overnight cultures of the indicated strains were sub cultured 1:100 and grown at 37°C in LB medium for 1 hour, then back diluted 1:100 in LB and incubated at 37°C for an additional 2 hours before imaging (C) MurA as well as the different mutants were purified, and their activity was measured in vitro by measuring the inorganic phosphate released from the MurA enzymatic reaction. (D) MurC and MurC^A132T were purified and their enzymatic activity was quantified in vitro by measuring the inorganic phosphate released from the MurC enzymatic reaction. (C-D) Statistical significance was determined by one-way ANOVA, error bars represent the standard deviation of three replicates (raw data points also shown).