Figure 4
From: Regulation of primate lentiviral RNA dimerization by structural entrapment
Effect of SL1 replacement in HIV-2 and SIV RNA chimeras. A. Schematic representation of HIV-2 and SIV SL1 with the substituted regions. HIV-2 and SIV sequences are represented with closed and open symbols, respectively. The SL1 sequences (nts 409 to 436) in 1–444 HIV-2 and 1–441 SIV (nts 410 to 433) RNAs were substituted by SIV and HIV-2 SL1 sequences, respectively. The SL1 sequences substituted encompass the apical loop and the stems C1 and C2. B. All four RNAs were individually incubated at 55°C in buffer H for 2, 4, 6, 8, 10, and 12 minutes before loading on a TBE/26°C agarose gel. A typical kinetics experiment with 1–444 HIV-2 RNA bearing wild type HIV-2 (lanes 1–7) or substituted SIV SL1 (lanes 8 to 14) is shown. Lane 1 represents monomeric RNA that was denatured at 90°C, then quenched on ice immediately prior to loading. C. Plots of the kinetic data for 1–444 HIV-2 (left panel) and 1–441 SIV (right panel) RNAs with wild type (open circles) or substituted SL1 (closed diamonds). The dimerization rate was extrapolated from the kinetics experiments according to [32]. The dimerization rate equals the slope of each linear curve divided by two (see Methods). For 1–444 HIV-2 RNA bearing wild type HIV-2 or substituted SIV SL1, kdim equals 0.11 ± 0.024 and 0.05 ± 0.007 μM-1min-1, respectively (duplicate experiments). For 1–441 SIV RNA bearing wild type SIV or substituted HIV-2 SL1, kdim equals 0.013 ± 0.004 and 0.026 ± 0.007 μM-1min-1, respectively (duplicate experiments).