Figure 6.

Direct binding of ANAC017 to a bipartite octamer predicted for H₂O₂ response. (A) The predicted target site of ANAC017 in an ATERF71 (AT2G47520) promoter was used as a biotinylated DNA probe in a DNA–protein binding assay by AlphaScreen. Underlining shows the predicted target site identified by the bipartite analysis. The sequences of unbiotinylated competitor DNAs with base substitutions from the biotinylated probe are shown (M0–M8, unsubstituted bases are shown as dots). Asterisks on the top of the probe sequence indicate critical bases for binding by ANAC017 protein, revealed by Panel B. (B) Results of the AlphaScreen assay are shown as the relative AlphaScreen signal (the signal obtained with non-biotinylated probe/signal with biotinylated probe). The average and standard error of four assays are shown.