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. 2022 Sep 8;10(5):e02311-22. doi: 10.1128/spectrum.02311-22

FIG 5.

FIG 5

Characterization of wild-type strains overexpressing mraZ, ΔmtrAB strains silencing mraZ at pH 10 stress, and binding of MtrA to the mraZ promoter analyzed by EMSA. (A and F) Growth curves of the Dietzia sp. DQ12-45-1b wild-type strain containing the mraZ overexpression plasmid and the ΔmtrAB strain containing antisense mRNA of mraZ plasmid from the acetamide-inducible promoter at pH 10. (B and G) Fluorescence microscopy of septation in wild type/pJV-mraZ (wild/pJV-mraZ) and ΔmtrAB/pJV-mraZ-AS under pH 10 stress. The branched phenotype is marked with arrows. (C, D, H, and I) Distribution curves and stacked columns of the percentage of cells containing a different number of septa from panels B and G were analyzed (~200 cells/sample). (E) EMSA analysis of MtrA, MtrA-P, MtrAD56E, and MtrAD56N binding to the mraZ promoter. MtrA, MtrAD56E, MtrAD56N, and MtrA~P at 0.5, 1, 2.5, and 5 μM were incubated for 30 min with 10 pM DIG-labeled Pmraz. (J) Proposed mechanism’s diagram illustrating the relationship between MtrAB TCS and the dcw cluster in Dietzia sp. DQ12-45-1b.