Chromosomal rearrangement by transposition.

<p>Chromosomal map of the 4 MDs (Ori: green, Right: red, Ter: light blue, Left: Dark blue) and the two non-structured regions (dashed black line) in the WT strain, the Right transposed strain and the Left transposed strain. <i>att</i> sequences used for the RT transposition are indicated by the green barre, the Right fragment between <i>attR127</i> (651 kb) and <i>attLlc13</i> (1099 kb)(dashed red line) was transposed at the position <i>attB O-NSR</i> (153 kb). <i>att</i> sequences used for the LT transposition are marked in blue, the NSL fragment between <i>attR124</i> (2892 kb) and <i>attL146</i> (3697 kb, dashed blue line) was transposed at the position <i>attB LT2</i> (1920 kb).</p

The position of the replication origin changes the Right and Left boundaries.

<p>Histograms of recombination frequencies between <i>att</i>L and <i>att</i>R sequences in WT, and Inv strain. Recombination frequencies between different <i>attR/L</i> sites are indicated on the y-axis. These values are the average of at least 3 independent experiments and the error bars correspond to the standard deviation. For both panels, strains were grown in minimal medium and the recombinase production was obtained by shifting cultures at 38°C for 10’. Relative position of each <i>att</i> sequence used in the experiment is represented on the MD map on top of each panel. The name of the <i>att</i> sequence, and the MD which they belong to, are indicated on the x-axis. The genetic backgrounds are indicated below the histogram (A), or with a color code (B). * indicates that the recombination frequency obtained was repeatedly 0.</p

The interaction limit of the Ori MD.

<p>Graphical representation of the Ori MD (green box), NSR region (gray line) and Right MD (red box). Coordinates of the <i>attR/L</i> sequences are indicated in fonction of their distance from the zero pb reference of the MG1655 genome for both configuration WT and RT. Percentages of recombination between <i>attL</i> and <i>attR</i> obtained after induction of 20’ at 36°C or 10’ at 37°C are shown. The histograms correspond to an average of at least 3 independent experiments with standard-deviations.</p

Position of chromosomal loci in the Ori and Right MD depending of <i>oriC</i> position.

<p>MD map of the <i>E</i>. <i>coli</i> chromosome are represented with the position of the Ori-4, Right-2, FROS-Tag in an Inv (A), or <i>oriCZ</i> (B) configurations. For each panel, the position of foci in cell containing 2 foci (cell with one focus are show in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1006758#pgen.1006758.s004" target="_blank">S4 Fig</a>), are represent in function of the long axis of the cell (y-axis) and in function of the cell length (x-axis). Red dots and bars correspond to the localization of the Right-2 loci and green ones to the Ori-4 loci. (C) Histogram of the percentage of the population presenting different interfocal distance (<0.2 to > 0.5) between Ori-4 and Right-2 in the WT strain (blue), Inv strain (red) and <i>oriCZ</i> strain (green). Numbers on the first columns show the percentage of cells where both foci are co-localized.</p

Position of chromosomal loci in the Ori, NSR region and Right MD.

<p>MD maps of the <i>E</i>. <i>coli</i> chromosome are represented with the position of the Ori-4, Right-2, NSR-5 FROS-Tag in a WT strain. For each panel, the position of foci in cell containing 2 foci, are represented in function of the long axis of the cell (y-axis) and in function of the cell length (x-axis). Green dots and bars show the position of the Ori-4 loci and red dots and bars correspond to the localization of the Right-2 (A) or NSR-5(B) loci. (C) Histograms of the percentage of the population presenting different interfocal distance (<0.2 to > 0.5) between Ori-4 and Right-2 (dark blue) or Ori-4 and NSR-5 (light blue).</p

Effect of Right and Left transposition on long distance DNA interactions.

<p>Histograms of recombination frequency between <i>attL</i> and <i>attR</i> sequences in WT, RT, and LT strains. The y-axis indicates the percentage of recombination between <i>attL</i> and <i>attR</i> sequences, obtained as described in Materials and Methods with 20’ induction at 36°C. The relative position of each <i>att</i> sequence used in the experiment is represented on the MD map on top of each panel. Histograms show the average of at least 3 independent experiments with their respective standard-deviation. Frequencies obtained in the WT strain are represented with the black bars, in the RT strain (A,C) or LT strain (B,D) with the gray bars and in the Δ<i>matP</i> RT strain (C), or Δ<i>matP</i> LT strain (D) with the light gray bars. * indicates that no <i>att</i>L<i>-att</i>R recombinant was obtained.</p

Extensive chromosomal disorganization in mutants of the ParABS system.

<p>(A) Percentages of the population presenting a given number of foci corresponding to the Ter locus or the Ori locus. Positioning of chromosomal loci located in the Ori region (left panels) and in the Ter region (right panels) in a wild type PAO1 strain (B), the Δ<i>parA</i> mutant (C) and the Δ<i>parB</i> mutant (D) grown in minimal medium supplemented with citrate. The position of the foci in cells containing 1 (upper panels) or 2 (bottom panels) foci are presented. Ori locus: 82-R in (A), (B) and (C). Ter loci: 2,957-R in (A), 3,090-L in (B) and (C).</p

Progressive segregation of <i>P. aeruginosa</i> chromosomal loci.

<p>(A) Position of each chromosomal locus on the PAO1 chromosomal map. Each locus is represented by a colored tag on the PAO1 chromosome according to its position. The color code is used for all figures. Black tags indicate rRNA operons. Grey tags represent <i>parS</i> sequence positions. Four of these <i>parS</i> sites are clustered close to <i>oriC</i>, four more are dispersed between positions 851-L and 628-R, and two are localized in the “right” replichore. (B) Average percentage of one-focus cells (solid diamonds) and two-foci cells (open diamonds) in a bacterial population grown in minimal medium supplemented with citrate. Cells with no visible focus were removed from this analysis (they constituted between 5 to 10% of the cells). The proportion of cells exhibiting more than two foci was always smaller than 0.5 %. The <i>x</i>-axis represents the positions of the loci on the chromosomal map. Values from four to eleven experiments were averaged, and the error bars represent standard deviations. (C) The average percentage of two-foci cells in a bacterial population grown in minimal medium supplemented with citrate, according to cell size, for each chromosomal locus of the “right” replichore (left panel) or of the “left” replichore (right panel). Values from four to eleven experiments were averaged, and the error bars represent standard deviations.</p

Proposed model for <i>P. aeruginosa</i> chromosomal organization.

<p>Organization in minimal medium supplemented with citrate (A) or glucose and casamino acids (B). Black lines represent fully replicated chromosomes, whereas grey lines represent partially replicated chromosomes. Colored markers represent chromosomal loci, and yellow diamonds represent replisomes.</p

Localization of <i>P.aeruginosa</i> DNA polymerase.

<p>EGFP-labeled replisome protein DnaX was observed in minimal medium supplemented with citrate (A) or glucose and casamino acids (B). For each panel, the upper left area shows a sample of representative cells; the lower left area presents the amount of cells exhibiting zero (white), one (blue), two (red) or three (green) fluorescent foci according to cell size. The upper right area presents the relative positions of the focus in one-focus cells, and the lower left area presents the relative positions of the foci in two-foci cells.</p