Fig. 6. Mapping of the KIF13B binding site within centaurin-₁ using yeast two-hybrid analysis. (A) Schematic representation of the domain structure (GAP domain, aa 1-126; N-PH, aa 130-230; C-PH, aa 253-356) and various deletion mutants of centaurin-₁. (B) Interaction of KIF13B with the full-length (FL) and deletion constructs of centaurin-₁. The full-length and deletion mutants of centaurin-₁ fused to the DNA binding domain of LexA (bait) pBTM, were co-transformed with KIF13B fused to the activation domain of Gal4 (prey) into the L40 yeast strain, and the transformants analysed for their ability to grow on medium lacking histidine, and to metabolise X-gal by ß-galactosidase. Panels, left to right, show three clones of each transformant grown on medium with (+H) and without (–H) histidine, and the filter ß-galactosidase assay (ß-gal). The experiment was performed three times with three different yeast transformations with identical results.