Figure 1
From: ATPase activity tightly regulates RecA nucleofilaments to promote homologous recombination
CE-LIFP and EM analysis of the RecA-saturated nucleofilaments stimulated by the non-hydrolyzable ATP analog ATPγS. (a) Typical electropherograms of CE-LIFP analysis of RecA assembly on ssDNA (TMR-ss83mer) in the presence of ATPγS (0.1 mM) and Mg2+ (10 mM). The reaction solution included 3.0 μM RecA, 10 nM TMR-ss83mer and 0.1 mM ATPγS. Iv and Ih represent the intensities of vertically and horizontally polarized fluorescence, respectively. (b) EM of the RecA-saturated nucleofilaments stimulated by ATPγS. The reaction solution included 3.0 μM RecA, 300 nM ss83mer and 0.1 mM ATPγS. (c) CE-LIFP electropherograms for analysis of the RecA-saturated nucleofilaments in the presence of 0.1 mM ATPγS but absence of Mg2+. The reactions also contained 10 nM TMR-ss83mer and 3.0 μM RecA protein, and proceeded at 37 °C for 10 min. IS indicates internal migration marker (a, c). Peak 1: RecA-saturated nucleofilaments; Peak 2: unbound TMR-ss83mer. Note: (1) In all figures, the concentrations of the oligomers are given in terms of whole oligomer molecules; (2) except in Figure 1a, only Ih signals are displayed in CE-LIFP electrophoregrams; (3) IS indicates internal migration marker in all figures.
