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. 2023 Dec 14;14:8339. doi: 10.1038/s41467-023-43823-w

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© The Author(s) 2023

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — A Schematic representation of the B. subtilis replication origin and BUS sequence elements. DnaA-boxes are indicated with triangles. B Linear arrangement of DnaA domains I-IV. Locations of critical activities are shown. Endpoints of DnaA proteins used for various experiments are indicated. C Schematic describing single molecule TIRF microscopy experiments to investigate BUS activity. BUS sequence elements are indicated by colour. D Fields of view from single molecule TIRF experiments performed using different conditions (indicated on top) as a function of time. Red circles indicate spots that disappear over time. E Graph representing the percentage of fluorescent signals with decreased intensity over time in the presence of either ATP or ADP. F Graph reporting the percentage of fluorescent signals with decreased intensity over time using wild-type or mutant scaffolds. G Graph representing the percentage of fluorescent signals with decreased intensity over time using DnaA variants. DnaA^104–446 lacks domains I-II. DnaA^R264A is defective in oligomerisation. DnaA^I190A is defective in ssDNA binding. For all graphs, data shows the average and percentage standard deviation from three independent experiments (source data are provided as a Source Data file).