Interestingly, the cells harbouring the two AidB-YFP foci are significantly (p < 0.005) smaller
Fludarabine (1.93 μm on average) than the bacteria having a single focus of AidB-YFP at the constriction site (2.08 μm on average), suggesting that in the cell cycle, bacteria with 2 foci precede those with a single focus at the constriction site (Figure 3A). This feature of the cell cycle is depicted in the discussion. Figure 3 Size distribution of B. abortus carrying AidB-YFP, in the presence or absence of an alkylating agent (EMS). The bacterial lengths were grouped in classes of 0.25 μm, and the maximum value for each class is given on the × axis. (A) Size distribution of 276 bacteria (XDB1128 strain) with AidB-YFP either at the new pole (white), the new pole and the constriction site (dark grey), or the constriction site only (black). (B) Size distribution of B. abortus (XDB1128 strain) exposed to 0.4% of EMS for 4 h (light grey, n = 340) or the unexposed control (white, n = 218, bacteria without detectable constriction). selleck inhibitor (C) DIC and fluorescence pictures of the XDB1128 strain expressing aidB-yfp and pdhS-mCherry fusions, as described in figure 2. The bacteria in the lower panels have been exposed to 0.4% EMS for 4 h in rich (2YT) medium. On the
top panels, control bacteria were incubated for 4 h in 2YT in the absence of EMS. Constriction sites are indicated by arrowheads. Each scale bar represents 2 μm. Furthermore, the localization of AidB-YFP is still at the new pole after 4 h of exposure with 0.4% EMS (80% of the bacteria exhibited PdhS-mCherry at one pole and AidB-YFP at the opposite pole, n = 237). This selleck screening library observation indicated that AidB-YFP is not released from the new pole in the presence of an alkylating stress with EMS, further suggesting that AidB is active at the new pole, because in these conditions an aidB mutant is killed. Interestingly, bacteria exposed to EMS displayed detectable constriction at the much less frequency (2 constrictions observed among 254 bacteria) compared to the
untreated control (44 constrictions observed among 254 bacteria). Moreover, bacteria treated with 0.4% EMS for 4 h and Etofibrate were significantly (p < 0.001) longer on average than unconstricted bacteria that were not exposed to EMS (Figure 3B). This suggests that growth is not arrested by the presence of EMS, while constriction is clearly inhibited. This is consistent with a replication arrest caused by alkylation of the bacterial genome, as previously reported for E. coli . AidB polar localization persists inside host cells B. abortus is an intracellular pathogen that encounters various stresses during its life cycle . Since these stresses could result in the alkylation of DNA, e.g. through nitrosative stress , we tested the localization pattern of AidB-YFP in B. abortus (XDB1120 strain) during an infection of human epithelial cells (HeLa cells).