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Fig. 5. FBF binds specifically to elements in fog-1 and fog-3
3'UTRs. (A) Putative FBF binding elements (FBEs) in fog and
fem 3'UTRs; black triangles, elements that bind in vitro; gray
triangles, elements that do not bind in vitro. (B) Nucleotide sequence of
predicted FBF binding elements. Sequences are aligned by conserved UGU motif
(bold). Mutated nucleotides are lowercase. [UUUUU towards 3' end of
fog-1 FBE bc RNA was disrupted to increase expression in three-hybrid
system (indicated by `a').] (C) Yeast three-hybrid assay. (D) Yeast
three-hybrid results. Error bars represent standard deviation of at least four
repetitions. gld-1 FBEa, element in gld-1 3'UTR
(Bernstein et al., 2005;
Crittenden et al., 2002);
fem-3 PME, element in fem-3 3'UTR
(Zhang et al., 1997), which is
called fem-3 FBEa in A. (E) FBF-2 binds FBEs in fog-1 and
fog-3 3'UTRs. Protein concentrations were 0.5 nM, 1 nM and 5
nM, then consecutively doubled to reach 640 nM. Apparent binding constants are
shown below each shift.
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