1. EMSA Shows MYB80 Is Able to Bind to the Promoter of All Three Target Genes at Several Different MYB Binding cis-Elements.(A) The UNDEAD probe contains three MYB binding cis-elements.
EMSA Shows MYB80 Is Able to Bind to the Promoter of All Three Target Genes at Several Different MYB Binding cis-Elements.(A) The UNDEAD probe contains three MYB binding cis-elements. Underlined sequences represent unlabeled competitors “a” and “b.”(B) The VGD1 probe contains two MYB binding cis-elements with a single unlabeled competitor “c” underlined.(C) The GLOX1 probe contains two MYB binding cis-elements. Underlined sequences are unlabeled competitors “d” and “e.” Numbers denote location relative to the ATG translational start site. Highlighted colors represent the motifs MYB1AT (green), MYB1LEPR (yellow), and MYBPZM (blue).(D) MYB80 is able to bind to all three cis-elements present in the UNDEAD promoter. Nonlabeled competitors are able to reduce the visible shift significantly (arrows), resulting in an increase in free probe.(E) MYB80 is able to bind to both MYB1AT cis-elements present in the VGD1 promoter. The unlabeled “c” competitor even at 100-fold does not significantly reduce the second visible shift, suggesting GAAACCA is not the preferred motif. The reduction is more prominent in the supershift using a T7 antibody against the fusion MYB80 protein.(F) MYB80 preferentially binds to the MYBPZM cis-element (CCAACC) in the GLOX1 promoter as “d” is the most effective competitor at reducing the visible shifts. neg, free labeled probe (no MYB80 protein); pos, labeled probe and MYB80; a-e/25/100, pos + unlabeled competitors at 25- and 100-fold compared with labeled probe.
Huy Anh Phan et al. Plant Cell 2011;23:
©2011 by American Society of Plant Biologists