Figure 1.
JAZ Proteins Differentially Interact with COI1 in the Presence of JA-Ile.
(A) JA-Ile differentially promotes COI1 interaction with various JAZ proteins. Pull-down assays were performed using crude leaf extracts from 35S:COI1-Myc transgenic plants and purified recombinant JAZ-His proteins. Reactions were supplemented (+) with 1 μM (3R,7S)-JA-Ile or an equivalent volume of assay buffer (−). Protein bound to JAZ-His was separated by SDS-PAGE and analyzed by immunoblotting with anti-Myc antibody for the presence of COI1-Myc. The blotted membrane was stained with Coomassie blue to visualize the amount of JAZ-His loaded.
(B) JAZ8 does not associate with COI1 in the presence of JA-Ile. Pull-down assays with purified JAZ8-His and JAZ10.1-His were performed as described in (A). Reaction mixtures were supplemented with the indicated concentration of (3R,7S)-JA-Ile.
(C) JAZ8 weakly associates with COI1 in the presence of coronatine. Pull-down assays were performed as described in (A). Reactions were supplemented with the indicated concentration of coronatine (COR).
(D) Saturation binding of 3H-labeled coronatine to COI1-JAZ8 (circle) and COI1-JAZ10.1 (triangle) complexes. Data show the mean ± se of two replicates performed in duplicate. CPM, counts per minute.