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. 2021 Mar 12;17(3):e1009452. doi: 10.1371/journal.pgen.1009452

Fig 11. Yeast 2-hybrid assays identify distinct Dis2 and Swd22 binding sites in Ppn1.

Fig 11

(A) Full-length Ppn1 (aa 1–710) and the indicated Ppn1 fragments fused to the Gal4 AD were tested for 2-hybrid interactions with Gal4 BD fusions to full-length Dis2 and full-length Swd22. AD/BD pairs that scored positive in both reporter assays (LacZ expression and histidine prototrophy) are indicated as ++. Pairs that were negative in both reporter assays are scored as --. (B) Full-length AD-Ppn1 constructs with the indicated amino acid substitutions were tested for 2-hybrid interactions with BD-Dis2 and BD-Swd22. (C) The amino acid sequences of the PP1/Dis2 binding motifs in PNUTS and Ppn1 are aligned. The conserved hydrophobic amino acids important for the Ppn1-Dis2 interaction in the 2-hybrid assay (Val, Trp, Leu) are in white font on black background. Conserved hydroxyamino acids that are potential phosphorylation sites are denoted by dots. The upstream basic patch is underlined.