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. 2020 Mar 20;11:1478. doi: 10.1038/s41467-020-15182-3

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a The wild-type uvrA allele was replaced by the mutant allele in mfd-YPet cells, so that mutant cells either express the distal ATPase mutant UvrA(K646A) or the proximal ATPase mutant UvrA(K37A) from the chromosome. b Bar plots show lifetimes of DNA-bound Mfd-YPet in the corresponding genetic backgrounds. Where two kinetic sub-populations are detected, the fast lifetime is displayed in the lower panel. Numbers in percentages represent the amplitudes of kinetic sub-populations (see Supplementary Figs. 2, 3). c Lifetime of Mfd-YPet in mfd-YPet uvrA(K646A) ΔuvrB (26 ± 2 s; N_repeats = 6 totaling n_obs = 23,763 individual observations) is similar to that of Mfd-YPet in ΔuvrA cells (29 ± 2 s²¹), suggesting the distal ATPase mutant (gray) is unable to interact with Mfd (green). d Lifetime of Mfd-YPet in the presence of UvrA(K646A) and UvrB (orange) (37 ± 3 s; N_repeats = 11 totaling n_obs = 36,073 individual observations) also resembles that of Mfd-YPet in ΔuvrA cells. e Mfd-YPet is arrested in the presence of the proximal ATPase mutant UvrA(K37A) in cells lacking UvrB (lifetime of 304 ± 69 s; N_repeats = 7 totaling n_obs = 45,254 individual observations). f Mfd-YPet dissociates in 52 ± 4 s (N_repeats = 8 totaling n_obs = 36,486 individual observations) in the presence of UvrA(K37A) and UvrB, suggesting that ATP hydrolysis at the proximal site is required for promoting the dissociation of Mfd-YPet. Error bars are standard deviations from ten bootstrapped CRTDs. Source data are provided as a Source Data file.