Extended Data Figure 6 : Lifetime distributions of the Mfd–RNAP intermediate as a function of UvrA concentration, using the tethered-DNA assay.

From: Reconstruction of bacterial transcription-coupled repair at single-molecule resolution

Extended Data Figure 6

The DNA substrate used in these experiments was positively supercoiled and contained a C-less cassette, and data were collected using the continuous-tracking methodology in the presence of 10–20 pM RNAP holoenzyme, 500 nM GreB, 100 nM Mfd, 2 mM ATP, 200 μM UTP, 200 μM GTP and 250 nM UvrB. The UvrA concentration was (a) 50 pM, (b) 75 pM and (c) 100 pM. Red lines show the result of global fitting to a difference-of-two exponentials characteristic of a Michaelis–Menten process, using the rate-limiting forward catalytic step extracted from classical Michaelian analysis of the mean times (Fig. 2d) as an additional constraint. d, Overview of lifetimes of the Mfd–RNAP complex measured with the tethered-DNA assay and as a function of template supercoiling, cause of RNAP stalling and UvrA concentration, as presented in this paper. UvrB was fixed at 250 nM throughout.