Journal: eLife

Article Title: Single-molecule visualization of fast polymerase turnover in the bacterial replisome

doi: 10.7554/eLife.23932

Figure Lengend Snippet: Individual intensity trajectories for 300 ε and τ foci were simulated in MATLAB 2014b. The simulation allows us to set k on and k off (in units of frames –1 ) for ε exchanging into Pol III*, and k on and k off for Pol III* exchanging into the replisome. By changing these rate constants, we can simulate different exchange mechanisms. The black line represents the average cross-correlation function for Pol III* exchange (both ε and τ). Here, k on and k off for Pol III* were set to 0.01 and the rate constants for ε were set to k on = 1 and k off <<1 to simulate stable binding of core to Pol III*. A clear positive peak can be seen. The green line represents the average cross-correlation function for simulated trajectories without any exchange. Here, k on = 1 and k off <<1 for all rate constants, to simulate stable binding to the replisome. In this case, there is no positive cross correlation. The gray line represents the average cross-correlation function for core exchange. In this case k on = 1 and k off <<1 for Pol III* and k on and k off for ε were set to 0.01. Again, we do not observe a positive cross correlation. Cross-correlation functions have been vertically offset for clarity. DOI: http://dx.doi.org/10.7554/eLife.23932.024

Article Snippet: The histograms obtained were fit with a Gaussian distribution function using MATLAB 2014b, to give a mean intensity of 5100 ± 2000 for the red and 1600 ± 700 for the green Pol III core ( ).

Techniques: Binding Assay