Figure 4

Identification of TBP/HAP1 interaction domains. The interaction of truncated HAP1 clones with TBP-C, TBP-N, and auto-activation controls are shown. Yeast containing the indicated bait and prey plasmids (listed below) were grown on SC-L-W (top panel), which selects for the presence of the bait and prey plasmids, and SC-L-W-H + 50 mM 3-AT (bottom panel), which selects for the interaction between bait and prey. All baits were expressed from the pDBLeu plasmid and all truncated HAP1 preys were expressed from the pPC86 plasmid. Sectors 3, 6, 9, 12, 15, 18, 21, 24, 27, 30, 33, 36, and 39 were controls confirming that truncated HAP1 clones were not auto-active. Sectors 2, 5, 8, 11, 14, 17, 20, 23, 26, 29, 32, 35, and 38 confirmed that none of the HAP1 clones interacted with TBP-N. Clones 10, 13, 19, and 22 did not grow on SC-L-W-H + 50 mM 3-AT, which suggested that these clones lacked the TBP interaction domain, whereas growth of clones 1, 4, 7, 16, 25, 28, 31, 34, and 37 on SC-L-W-H + 50 mM 3-AT suggested that these clones contained a TBP interaction domain. Three independent clones of all HAP1 truncations were tested and each gave the same result; one representative set is shown.