Multiple chloroplast proteins bind to different regions of the atpI 5'UTR in vitro
Band shift assays with radiolabeled full-length atpI 5'UTR were performed in the presence (+) or absence (-) of unlabeled RNAs corresponding to either the 5' or 3' regions of the 5'UTR. Diagrams to the right of the images denote the migration location of full-length 5'UTR with one or both of two different proteins bound
Band shift assays were transferred to membrane filters and probed with antibodies against chloroplast ribonucleoproteins (cpRNPs)
cpRNP antibodies were a generous gift of ProfessorMasahiro Sugiura and colleagues, Nagoya Japan
There are at least two different proteins that bind the atpI 5'UTR
cpRNPs are present in atpI 5'UTR-protein complexes in vitro
+
-
+
+
+
+
+
-
-
+
+
+
+
+
+
-
-
-
radiolabeled wild-type
(unlabeled) 5'partial
protein
radiolabeled wild-type
3'partial (unlabeled)
protein
* atpI 5'UTR-protein complexes
*
*
Antibodies against:
atpI 5'UTR:
cp29A/B
cp28/31
cp33
-
+
-
+
-
+
wild-type:
partials:
-155
-111
-96
-57
-22
-10
-1
+15
-80
-80
-1
5'partial
3'partial
Con1
Con2
RBS
ORF
-155