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Figure 3


Fig. 3. Mapping of the LFY interaction domain. (A) Diagram of the mutated and truncated forms of LFY. Gray boxes indicate the conserved domains at the N terminus and C terminus. Either D139 or S140 were mutated to alanine and used for yeast two-hybrid assays. Four different truncated versions of LFY, LFYN1 (amino acids 1-141), LFYC1 (amino acids 142-420), LFYN2 (amino acids 1-375) and LFYC2 (amino acids 376-420) are shown. (B) Yeast two-hybrid assays testing the interaction between various LFY truncations/mutants and {Delta}FUFO. Quantitative measurements of β-GAL activities (average of five independent assays) are indicated. LFYN2 fails to interact with {Delta}FUFO, while LFYC1 retains the interaction. Neither of the mutant forms D139 nor S140 affected the interaction with {Delta}FUFO. Bars represent mean±s.e.m. for the five replicates. (C) GST pull-down assays performed with bacterially expressed GST fusion proteins and inflorescence protein extracts from 35S::UFO-Myc plants. Top panel, western blot probed with anti-Myc antibody. Bottom panel, Coomassie Blue stained gel showing GST fusion proteins used in the assays. LFYN2 does not pull down UFO as efficiently as does full-length LFY from plant extracts.





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