Fig. 5: Specific interactions of MraZ subunits with the DNA binding boxes.

a Cartoon representation of the complex between four binding DNA boxes with four MraZ subunits. DNA boxes and subunits are labeled and shown in different colors. b Surface representation four DNA binding boxes in complex with four MraZ subunits. c Schematic representation of the interactions between side chains and nucleobases of the four binding boxes. Interactions are shown by short lines. d Close up view of the contacts between Lys13, Arg15 and Arg86 with the consensus sequence of the binding box in the DNA major groove. Contact residues with nucleobases and distances are labeled and shown in stick representation. Sharpened volumes are displayed for the contact region.e Opposite view of (d). f Agarose gels of EMSA experiments of MraZ wild type and K13A, R15A and R86A with fluorescent dsDNA containing four boxes. g Repression assays containing GFP-based transcriptional reporter under the control of MraZ promoter binding boxes with overexpression of WT or DNA-binding mutant MraZ protein. Data values represent the mean ± SEM, n = 3 technical replicates. Significance was measured by a two-tailed unpaired t test relative to the positive GFP expression control. All data were analyzed with a 95% confidence interval. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001. Exact P values from left to right: 0.0001 and 0.0026. h Bar plot of the RT-qPCR experiment showing the GFP mRNA levels of a GFP-based transcriptional reporter under the control of the MraZ promoter with overexpression of WT or DNA-binding mutant. Data values represent the mean ± SEM, n = 3 technical replicates. Significance was measured by one-way ANOVA with Tukey post-hoc test. Exact P-values from left to right: 0.0004 and 0.1535.