(A) EMSA gels showing
binding of full-length (ZF1–4) and
two-finger (ZF12, ZF23, and ZF34) JAZ constructs to full-length VAI
RNA. In each gel, the first lane is free RNA. In the second lane,
the protein concentration is 20 nM (4 nM for ZF34). In each subsequent
lane, the protein concentration is doubled until [JAZ] = 5 μM
(1 μM for ZF34) in the tenth lane. (B) EMSA gels showing binding
of JAZ ZF1–4, ZF12, ZF23, and ZF34 to T57A RNA. In each gel,
the first lane is free RNA. In the second lane, the protein concentration
is 20 nM (4 nM for ZF34 and ZF23). In each subsequent lane, the protein
concentration is doubled until [JAZ] = 5 μM (1 μM for
ZF34 and ZF23) in the tenth lane. The figures of the RNA structures
were produced using Mfold.38 Full sequences
with RNA bases shown in larger type are given in Supporting InformationFigures S2 and
S3. The JAZ constructs are identified with colors corresponding
to those in Figure 1B.