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. 2019 Jun 27;38(15):e101649. doi: 10.15252/embj.2019101649

Figure 6. DnaA‐boxes and DnaA‐trios are required for specific DNA unwinding by DnaA.

Figure 6

  1. Schematic of the DNA unwinding assay using an oligonucleotide scaffold.
  2. DNA unwinding requires DnaA‐boxes and DnaA‐trios.
  3. Schematic of the electrophoretic mobility shift assay using an oligonucleotide scaffold.
  4. Electrophoretic mobility shift assay showing that DnaA‐boxes are required to form a stable complex with DnaA. ADP was present at 1 mM, and DnaA concentration was 0, 63, 125, 250 nM.
  5. Schematic of the DnaACC filament assembly assay using a DNA scaffold and the cysteine‐specific crosslinker BMOE.
  6. DnaACC filament assembly on the DNA scaffold requires DnaA‐trios.
  7. Schematic of the DNA unwinding assay using oligonucleotide scaffolds with either one (ΔA) or three (ΔATG) base pair deletions.
  8. The rate of DNA unwinding decreases when one base pair is deleted but is similar to wild‐type when three base pairs are deleted.