FIGURE 15.

Two-hybrid assessment of Tor1 association with full-length Gln3(1–730) (pRR1065), Gln3(508–730) (pRR1248), and Gln3(508–730)_{S656D,S659D,S662D} (pRR1250). The assays were performed as described under “Materials and Methods.” The transformation recipient in all cases was strain PJ69-4a. The positive control was constructed by sequentially transforming PJ69-4a with pAV3-1 and pTD1-1. The negative control was a transformant containing pASTOR1-2470 or pASTOR1-1764 (Gal4 DNA binding domain-Tor1 fusion) and pACT2 (corresponding empty vector). pRR1065, pRR1248, and pRR1252 consisted of the Gal4 activation domain fused N-terminal of the respective gln3 genes. Viability and the test for a two-hybrid association of the transformants were determined by streaking them on the same medium in the presence and absence of 3 mm 3′-aminotriazole (3AT). Plates were incubated 78 h. The procedures, strain, fusion vectors, and control plasmids were those used by Carvalho and Zheng (23).