Figure 4.

The pex7-2 lesion disrupts PEX5 and PTS2-cargo binding in yeast. (A) Yeast two-hybrid assays were conducted with PEX7 and pex7-2 fused to the GAL4-DNA binding domain (BD) and PEX5, PED1 (a PTS2 protein), and tPED1 (without the PTS2 region) fused to the GAL4 activation domain (AD). Serial dilutions spotted onto permissive plates (+His) and plates on which growth requires interaction (−His) are shown after incubation at 30°C for 2 d. (B) Proteins extracted from yeast strains in A were separated using SDS-polyacrylamide gel electrophoresis and immunoblots were probed with the indicated antibodies. Extracts from 8-d-old wild-type Arabidopsis seedlings and the untransformed AH109 yeast were included as controls.