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. 2022 May 19;298(6):102051. doi: 10.1016/j.jbc.2022.102051

Figure 5.

Figure 5

Right-DnaA subcomplex binds DnaBC. A, biotinylated DOR fragments are shown by bars below the oriC structure. For symbols in the oriC structure, see Figure 1A. B, ATP–DnaA (10 pmol: 0.4 μM), His-DnaB (10 pmol as monomer), and DnaC (10 pmol) were incubated in the presence or absence (−) of biotinylated FL-, Left- or Right-DOR fragment (250 fmol, 10 nM) (Fig. 2A), followed by pull-down assay. Proteins bound to DNA were analyzed by SDS–11% PAGE and silver staining. +, presence; −. absence. C, amounts of recovered proteins (DnaA, DnaB, and DnaC) were determined using standard curves, and values from the negative control excluding the DOR were subtracted. About 150 fmol DNA was recovered. The means and SDs of the numbers of DnaA, DnaB, and DnaC molecules recovered per DNA are indicated by light-to-dark graybars, respectively. Five independent experiments were carried out, and the mean values with SDs (n = 5) are shown in the graphs with each data as corresponding color dots. DOR, DnaA oligomerization region; FL, full length.