Lead(II)-induced cleavage
patterns of M1 RNA in the presence of increasing concentrations
(in mM) of the Mg(H2O)62+ analog
Co(NH3)63+ and spermidine, as
indicated. Cleavage was performed at the indicated concentrations
and 37°C, as outlined in Materials and
Methods and Figure 2 legend, with the exception that the time of
incubation in the presence of Pb2+ was 7 instead
of 6 min. Ctrl 1, incubation in the presence of 50 mM Tris–HCl
pH 7.5, 100 mM NH4Cl, 10 mM Co(NH3)63+,
10 mM spermidine, 10 mM MgCl2 and without Pb2+;
Ctrl 2, as Ctrl 1 but incubation in the presence of 50 mM Tris–HCl
pH 7.5 and 100 mM NH4Cl (without Pb2+)
only; Mg2+, Pb2+ cleavage
in the presence of 50 mM Tris–HCl pH 7.5, 100 mM NH4Cl
and 5 mM MgCl2. Roman numerals indicate the cleavage sites
as shown in Figure 1.