FIGURE 2.

Mapping of Msg5 regions that mediate MAPK interaction. A, schematic of full-length and Msg5 fragments analyzed for MAPK interaction (left) and qualitative analysis of the two-hybrid interaction by growth analysis of diploid cells bearing plasmids that encode the indicated proteins or protein fragments (right). Before mating, pGBKT7-Msg5, pGBKT7, pGBKT7-Msg5(126–489), pGBKT7-Msg5(1–123), pGBKT7-Msg5(90–489), pGBKT7-Msg5(1–45), or pGBKT7-Msg5(46–489) were transformed into PJ69-4A cells, and pGADT7 and either pGADT7-Kss1, pGADT7-Fus3, pGADT7-Slt2, or pGADT7-Mlp1 were transformed into PJ69-4α cells. Two-hybrid assays were performed as indicated in Fig. 1A. B and C, semiquantitative analysis of the two-hybrid interaction between Msg5 or the different Msg5 fragments and the distinct MAPKs based on the level of induction of β-galactosidase. Experiments were performed in triplicate on the diploid cells described in A, with error bars representing S.D.