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. 2014 Feb 20;10(2):e1004136. doi: 10.1371/journal.pgen.1004136

Figure 2. Identification of the Ddc2 region required for Mec1 interaction and DNA damage response using the modified two-hybrid assay.

Figure 2

The two-hybrid tester ddc2Δ strain (KSC2077) carrying pBD-MEC1(2-2368) was transformed with pAD-DDC2, pAD-DDC2-ΔC, pAD-DDC2-ΔN1, pAD-DDC2-ΔN2, pAD-DDC2-ΔN3 or pAD-DDC2-S4. The coiled-coil domain (amino acid 68–138) is indicated as a gray bar and the KKRK sequence (amino acid 177–180) is shown as KR. The ddc2-S4 mutation carries two substitution mutations (K263E and H382Y). Transformants were streaked on an SD-Ura-Leu-His plate containing 1 mM AT, YEPD plate containing 1 mg/ml HU or 0.005% MMS. Interaction with Mec1 and DNA damage sensitivity were assessed by cell proliferation on the respective plates.