Figure 1

Constructs used for split ubiquitin assays. Bait protein fusion was made with the C-terminal part of ubiquitin (Cub) followed by artificial transcription factor LexA-VP16 at the C-terminus of DOP-2XL (1, 2). Prey protein fusions were constructed with the mutant N-terminal part of ubiquitin (NubG) either at the C terminus of GPA-14 (3) or at the N-terminus (4). If bait and prey interact NubG and Cub are forced into close proximity, resulting in the reconstitution of split-ubiquitin and release of LexA-VP16 transcriptional factor that leads to transcriptional readout, resulting in growth of yeast on selective medium and color development in a β-galactosidase assay.