Skip to main content
. Author manuscript; available in PMC: 2014 Feb 1.
Published in final edited form as: Virus Res. 2012 Sep 24;171(2):287–303. doi: 10.1016/j.virusres.2012.09.008

Figure 7. DNA synthesis by HIV-1 RT on circular ssDNA template in the presence of NCp7 and F-actin.

Figure 7

The polymerase reaction was carried out over 40 min. as in Fig. 6. Concentrations of primed-ssDNA circles, RT and NCp7 were, respectively, 5 nM, 100nM, 3.4 μM. F-actin filaments were added to the assay at a concentration close to 0.1 μM (monomer). NCp7 and primed-ssDNA circles were premixed for 4 min. at 37°C, followed by incubation with RT for 2 min., F-actin for 2 min. before addition of dNTPs (100 μM each) to start the reaction. a and b: controls without F-actin showing clusters of dismantling NCp7-ssDNA aggregates due to dsDNA extrusion. c, d and e: reactions in the presence of F-actin showing binding of the NCp7-RT-DNA complexes all along the F-actin filaments. 5 μl-aliquots of the assay were diluted 5-fold in the reaction buffer without DTT and next deposited onto the EM grid. The scale bars correspond to 0.5 μm.