Figure 2: MukBEQEF clusters form slowly.
From: MatP regulates the coordinated action of topoisomerase IV and MukBEF in chromosome segregation
(a) Wild-type MukB or MukBEQ were expressed from an arabinose-inducible promoter at the chromosomal mukB locus. Transcription from the smtA promoter was blocked by terminators T1–T2. Repletion of wild-type MukB (AU2064) (b) and MukBEQ (AU2089) (c) in ΔmukB cells. Two methods were used to monitor focus formation; the average intensity of the brightest MukE-mYPet pixel in cells, as well as manual counting of foci (n=460–1,000 cells). Brightest pixel data are means (±s.e.m) of three experiments. Steady-state data were from cells where MukB/MukBEQ was expressed from their normal promoters. See also Supplementary Fig. 3. (d) MukBEF foci colocalize with ter during MukE depletion. MukBEF focus position was followed before (0) and after depletion. Left: snapshot images of cells with labelled ter3 and MukB are shown for the indicated depletion times. The arrow shows colocalization of MukB with ter on MukE depletion. Right: % ori1 (Ab86) or ter3 (Ab227) foci associated with a MukB focus, in cells with MukBEF foci, were plotted for each time point. After 80 min of depletion, cells had only weak MukBEF foci. n≥300.
