FIGURE 3.
VS ribozyme cleavage in trans is concentration dependent. Approximately 1 nM [32P]-labeled A756C VS RNA was incubated with G638A VS RNA at concentrations between 0.1 and 10 μM, and reaction progress measured as a function of time (A). The measured extents of cleavage are plotted as a function of G638A VS RNA concentration. The data for each concentration were fitted simultaneously to two exponential functions, with the amplitudes for the fast and slow cleaving fractions treated as global parameters. G638A VS RNA concentrations: 0.1 μM, open circles; 0.3 μM, closed circles; 0.75 μM, open triangles, inverted; 1.0 μM, closed triangles, inverted; 1.75 μM, open diamonds; 3.0 μM, closed diamonds; 4.5 μM, open squares; 6.5 μM, closed squares; 8.5 μM, open triangles; 10 μM, closed triangles. (B) Plot of the fast rates from the average of two sets of kinetic data, fitted to the Michaelis–Menten Equation 1.