Figure 1.

Interaction between AtPirin1 and GPA1.

(A) Reconstitution of the yeast two-hybrid interaction between AtPirin1 and GPA1. The indicated combinations of the bait (pLexA-NLS) and prey (pACT) constructs were transformed into the yeast reporter strain. Transformants were examined for β-galactosidase activity in the presence of 5-bromo-4-chloro-β-d-galactoside (+X-gal) and for growth in the absence of His (−His).

(B) Coomassie blue staining of the GST-GPA1 fusion protein and GST (positions of the GST-GPA1 and GST proteins are indicated by arrows). MW, molecular mass.

(C) In vitro protein interaction between AtPirin1 and GPA1. GST-GPA1 was overexpressed in E. coli, immobilized on glutathione-Sepharose 4B beads, and incubated with ³⁵S-labeled AtPirin1 protein obtained by coupled in vitro transcription/translation. AtPirin1 binds to GST-GPA1 but not to GST alone.