Fig. 4: The effect of the ΔDARS1, ΔDARS2, ΔdatA triple deletion on the binding of DnaA and the DNA replication of E. coli.

A The dnaA::PAmCherry2.1 mutation was introduced in the chromosomal locus of dnaA in the E. coli Δ3D strain. B Measurements of DNA content (top) (see Supplementary Fig. 5A for a complete list of replicates) and cellular area (bottom) of either the wild-type or the mutant strain for cells grown in slow (n_WT = 1409 cells, n_Δ3D = 1056 cells), intermediate (n_WT = 790 cells, n_Δ3D = 469 cells) or fast growth regime (n_WT = 693 cells, n_Δ3D = 480 cells). For the bar graph, data are presented as mean values ± SD, n = 3 biologically independent replicates. P-values are calculated by a two-sided T test. Box plots indicate median (middle line), 25th and 75th percentile (box) and 5th and 95th percentile (whiskers) as well as outliers (single points). C The diffusion coefficient histograms of DnaA obtained from sptPALM of E. coli Δ3D cells growing either in slow, intermediate or fast growth regime. Histograms are fitted (black line) with a theoretical description of 250,000 particles moving between a free and a bound state. The number of cells and tracks per histogram is shown in the upper left corner. On the right, the kinetic rates describing the fitting are provided. D Average bound fraction of DnaA in each growth regime for either the wild-type E. coli MG1655 or the triple deletion mutant E. coli Δ3D strain. Data are presented as mean values ± SD, derived mathematically from kinetic rates in (C). P-values are calculated by a two-sided T test. E Trends of DnaA mobility throughout the cell cycle either in slow (n_WT = 958 cells, n_Δ3D = 664 cells), intermediate (n_WT = 688 cells, n_Δ3D = 659 cells) or fast growth regime (n_WT = 697 cells, n_Δ3D = 705 cells). The average diffusion coefficient of each cell is plotted against the area of the cell, with the black line indicating the rolling median of the population. The grey ribbons represent the interquartile range, whereas the dotted magenta lines represent the wild-type references. See Supplementary Fig. 5F for the area distribution of the cells. Source data are provided as a Source Data file.