Figure 1

Physical interactions of ATP sulfurylase with Cys3 and Cna1 by two-hybrid analysis. Images show representative colonies of S. cerevisiae expressing pairs of bait and prey fusion proteins, which are described at the top. + represents the positive and—the negative control provided by Match Maker kit (Clontech). YL019 is the strain code for S. cerevisiae Y2HGold carrying the plasmids that encodes the ATP sulfurylase (Met3) as bait and the catalytic subunit of calcineurin A deleted for the auto inhibition domain Cna1ΔC as prey; YL018: interaction of ATP sulfurylase as bait with Cys3 transcription factor as prey; YL026 and YL027: ATP sulfurylase deleted for the atypical leucine zipper (Met3ΔLZ) as bait and the catalytic subunit of calcineurin A deleted for the auto inhibition domain (Cna1ΔC) as prey; YL028 and YL029: ATP sulfurylase deleted for the leucine zipper (Met3ΔLZ) as bait and the Cys3 transcription factor as prey. Each line represents a different plate composition that test for different reporter genes; DDO = double drop out (minus triptophane and leucine), where no reporter gene is activated and tested; QDO = quadruple drop out (minus tryptophan, leucine, histidine and adenine), where two reporters are tested and QDO/X/A = quadruple drop out added with aurobasidin; and X-α-Gal, where all four reporters are tested. Plates were grown for 96 h at 30 °C, and photographed after this period.