Figure 2. Demonstrating the flexibility and scalability of the system by varying interaction domains.

(A) Variants of the different complementary interaction domains used. The constitutive interaction domains mPDZ, SH3 and SYNZIP are shown on the left; the ABA inducible ABI-PYL interaction domains appear on the right. (B) Comparison of YFP signal normalized by cell size from constructs bearing the indicated interaction domains along with either a functional (blue histograms) or non-functional (gray histograms) phosphodegron in yeast treated with 10 μM α-factor as in Figure 1C. The vertical dashed black lines on the histograms represent the medians of the populations with functional degrons whereas the solid black lines represent the median of the populations with non-functional degrons. (C) Median fluorescence – shaded regions cover the interquartile range – and population histograms of the YFP signal normalized to cell size from cells expressing the ABA inducible ABI-PYL interaction domains fused to Fus3 and YFP, respectively for a range of ABA concentrations. The raw time-course data corresponding to these endpoint observations can be found in Figure 2—figure supplement 1.

DOI: http://dx.doi.org/10.7554/eLife.15200.008

Figure 2—figure supplement 1. Time course characterization of different interaction domain variants post induction with α-factor.

In this plot each subplot is labeled with the name of the interaction domain pair that was used to target the MAPK Fus3 to degrade a YFP reporter in the yeast strain. These yeast strains were grown up to log phase from saturated cultures for 5 hrs and then induced with 10 μM α-factor at time 0 to activate the kinase. The dashed lines are uninduced controls. In the case of the ABI-PYL strain two additional induction conditions were assayed, namely with ABA (100 μM) and with ABA and α-factor. For these cultures the medium used to grow the yeast up to log phase had ABA in it. The fluorescence of these cultures was then assayed at regular intervals using flow cytometry. Raw data for two replicates performed on different days under identical conditions is shown.

DOI: http://dx.doi.org/10.7554/eLife.15200.009