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Table 2 Yeast Two-hybrid Interaction† of Parafibromin Fused to Gal4DBD with Full-length Proteins Fused to Gal4AD

From: The parafibromin tumor suppressor protein interacts with actin-binding proteins actinin-2 and actinin-3

 

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*

+

  1. †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).
  2. *Interaction was also positive with parafibromin fused to LexA-DBD (construct from [22]).