Figure 2.

Survey of the sites of interaction between the COOH-terminal domain of BP230 fused to GAL4-BD and IF proteins fused to GAL4-AD: + and - indicate growth or no growth, respectively, on selective media. The PJ69–4A yeast strain was cotransformed (triple transformation) with pACT2, pACT2-URA plasmids encoding IF proteins in fusion with GAL4-AD and pAS2–1 plasmids encoding BP230 fused at its NH₂-terminus with GAL4-DNA-BD. To test the interaction with either vimentin or monomeric keratins, triple transformations were performed with either pACT2 or pACT2-URA without insert. After selection on SC-LWUra agar plates, eight colonies of each transformation were arrayed in 96-well microtiter plates and then transferred onto agar SC-LWUra (positive control), SC-LWUra without adenine, and SC-LWUra without histidine and supplemented with 2 mM 3-amino 1,2,3-triazole. Growth was estimated after 5 d of incubation at 30°C. Transactivation controls were performed systematically for each construct with the opposite vectors without insert. # indicates no growth on medium lacking adenine. Each experiment was repeated at least twice.