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. 2023 May 18;83(10):1573–1587.e8. doi: 10.1016/j.molcel.2023.04.015

Figure 6.

Figure 6

Supercoiling accumulation reduces the Gal4 residence time on DNA

(A) Schematic of the C-terminal HaloTag at the endogenous GAL4 in haploid yeast (1n).

(B) Representative image of a yeast cell showing a single JFX650-labeled Gal4-HaloTag molecule (arrowhead). Scale bars, 2 μm.

(C) Survival probability distributions for Gal4 residence times in −OsTIR −IAA (323 cells, 706 tracks; 33,520 particles) and +OsTIR −IAA (244 cells, 1,378 tracks; 53,430 particles), and +OsTIR +IAA (107 cells, 187 tracks; 13,465 particles) cells.

(D) Average residence time of Gal4 calculated from trajectories with residence times >5 s for the indicated conditions. Errors indicate SEM, determined by bootstrapping with 300 repeats. ∗∗p < 0.01; ∗∗∗p < 0.001.

See also Figures S6 and S7 and Video S2.

Video S2. Example movie of Gal4 single molecules, related to Figure 6
A representative single-plane video of induced yeast cells expressing Gal4-HaloTag molecules which are sparsely labeled with JFX650 dye. The cells were imaged with 200 ms intervals for 900 frames
Download video file (4.3MB, mp4)