Table 2.
Yeast Two-hybrid Interaction† of Parafibromin Fused to Gal4DBD with Full-length Proteins Fused to Gal4AD
| Activation Domain Fusion | β-Galactosidase Assay | |
| 1 | Actinin-1 | - |
| 2 | Actinin-2* | ++ |
| 3 | Actinin-3* | +++ |
| 4 | Actinin-4 | - |
| 5 | α-actin | - |
| 6 | FHL-1 | - |
| 7 | FHL-2 | ++ |
| 8 | FHL-3 | - |
| 9 | FHL-4 | - |
| 10 | Desmin | ++ |
| 11 | β-catenin* | + |
†Yeast strain MaV203 that contains the Gal4-LacZ reporter or EGY48/pSH-18 (for β-catenin) that contains the LexA-LacZ reporter was transformed with plasmids expressing Gal4DBD- or LexA-DBD-fusion proteins together with plasmids expressing Gal4AD-fusion proteins. Transformed yeast colonies selected in SD medium lacking Leu and Trp (Leu, Trp and Ura for EGY48/pSH-18) were scored for β-galactosidase activity in colony filter lift assays. Interaction is indicated by β-galactosidase activity level. -, +, ++, or +++ indicate no, low, medium or high levels of β-galactosidase activity, respectively. None of the Gal4AD fusions interacted with Gal4DBD alone or with Gal4DBD-p53 (aa 72–390 of mouse p53). The Gal4DBD-parafibromin did not interact with Gal4AD alone or Gal4AD-TAg (aa 87–708 of SV-40 T-antigen).
*Interaction was also positive with parafibromin fused to LexA-DBD (construct from [22]).