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. 2011 Feb 28;156(1):106–116. doi: 10.1104/pp.110.170894

Figure 3.

Figure 3.

Analysis of the PYR1 mutations and their effect on the HAB1-dependent inhibition of OST1 activity. A, Interaction between HAB1 and PYR1 variants was analyzed by the Y2H growth assay on medium lacking His and Ade in the presence of 5, 10, or 20 μm (+)-ABA. Immunoblot analysis using antibody against the Gal4 binding domain (GBD) verifies the expression of the different fusion proteins in the Y2H assay. Ponceau staining from a representative yeast protein is shown as loading control. GAD, Gal4 activation domain. B, Relative inhibition of HAB1 activity by the different PYR1 variants in the presence of 8 μm ABA with respect to wild-type PYR1 (100%; sd was below 7%). C, OST1 in vitro kinase activity assay in the presence of HAB1, PYR1 wild type, and mutated versions, ΔCABF2 and 10 μm ABA, when indicated. The autoradiography shows the levels of autophosphorylation of OST1. D, Quantification of ΔCABF2 phosphorylation levels in the previous assay using the phosphoimager Image Gauge V.4.0. se measurements are shown (n = 3). [See online article for color version of this figure.]