Figure 6

ADAAD alters the conformation of primer 2 DNA.

(A) CD spectra for DNA alone in the absence and presence of K⁺. (B) CD spectra for primer 2 DNA (heat-cooled in the absence of K⁺) in the presence of ATP alone and ADAAD alone. (C) CD spectra for DNA alone and DNA incubated with 0.1 μM ADAAD and 0.5 mM ATP for 5 minutes at 37 °C. (D) Comparison of CD spectra of DNA alone, DNA with ADAAD and ATP incubated for 20 minutes at 37 °C before and after addition of 5 μM ADAADiN. ADAADiN was added after the DNA was incubated for 20 minutes in the presence of 0.1 μM ADAAD and 0.5 mM ATP at 37 °C. After addition, the CD spectra were recorded at 37 °C. As each scan takes 1 minute and 5 scans were taken, the reaction with ADAADiN had been effectively incubated for 5 minutes. (E) CD spectra of DNA (heat-cooled in the presence of 100 mM K⁺) alone, DNA with protein and ATP incubated at 37 °C for 5 minutes and after addition of 5 μM ADAADiN. (F) Comparison of CD spectra of DNA (heat-cooled in the absence and presence of 100 mM K⁺). In these reactions, 0.15 nM DNA, 0.1 μM ADAAD, 0.5 mM ATP and 10 mM MgCl₂ was used. ADAADiN is indicated as Inh. within the figure. In both cases, DNA was heated to 94 °C for 3 minutes and slowly cooled to room temperature either in the absence or presence of 100 mM K⁺.