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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. 4 — A Schematic representation of BHQ strand separation assay. The fluorescently labelled oligonucleotide contains an abasic site complementary to the central adenine (base shown in green). B Structures of nucleobases used to replace adenine. C Strand separation assay performed with nucleotide substitutions at the central adenine of the first DnaA-trio, in the context of an abasic site at the complementary position. Positions of mutations and structure of modified bases are indicated. Data shows the average and percentage standard deviation from three independent experiments (source data are provided as a Source Data file). D Schematic of the inducible repN/oriN system used to bypass mutations affecting oriC activity in B. subtilis. Replication via oriN is turned on and off in the presence and absence of IPTG, respectively. E Spot titre analysis with strains that have replaced the central adenine of DnaA-trio#1-3. The presence or absence of IPTG indicates the induction state of the repN/oriN system. Wild-type (FDS682), ΔincC (FDS404), A→G (FDS688), A→C (FDS686), A→T (FDS691). F Marker frequency analysis of strains shown in panel F. Data shows the average and individual data points from two independent experiments (source data are provided as a Source Data file). G Comparison of ssDNA substrates from DnaA nucleoprotein complexes. The synthetic polyA ssDNA from A. aeolicus (shown in green) (PDB 3R8F) was superimposed onto the BUS ssDNA (shown in yellow). Insets provide greater detail of the distinct conformations adopted by the two ssDNA substrates.