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Fig. 4. Determination of nos-2 3'UTR regions that are
critical for interaction with the various proteins. (A) Schematic
illustration of the five regions of nos-2 3'UTR that were
mutated by substitution. SubA begins immediately downstream of the stop codon.
(B-E) Electrophoretic mobility shifts of various mutant versions of
radiolabeled nos-2 3'UTR by MBP:MEX-3 (B), GST:SPN-4 (C) and
GST:OMA-2 (D,E). The first lane in each set is the mobility of RNA in the
absence of protein. Radiolabeled RNA used in D contained the wild-type version
of the following regions only: 1, SubA-C; 2, SubB-E; 3, SubA-D; 4, SubD-E,
whereas those in other panels contained the 200 bp nos-2 3'UTR
with the indicated regions substituted with (TG)15. WT in all
panels indicate the wild-type version of the 200 bp nos-2
3'UTR. (F) Binding of radiolabeled WT and mutant nos-2
3'UTR RNA to solid matrix in presence of the indicated components (see
Materials and methods for details). L RNA, radiolabeled RNA; UL RNA, unlabeled
RNA.
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