Interaction in vitro; bait produced by in vitro translation; prey produced by in vitro translation.

hpo homodimerizes.

Two-hybrid system: yeast

Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey from cDNA expression library).

Source was cell extract of S2 cell line; bait produced from transfected construct; prey produced from transfected construct.

The I634D mutation completely disrupts SARAH-mediated hpo homodimerization. However, full length hpo bearing the I634D mutation retains some homodimerization activity, suggesting that other regions also contribute to interaction.

Source was cell extract of S2 cell line; bait produced from transfected construct; prey produced from transfected construct.

The I634D mutation completely disrupts SARAH-mediated hpo homodimerization. However, full length hpo bearing the I634D mutation retains some homodimerization activity, suggesting that other regions also contribute to interaction.

Source was live S2 cells; bait produced from transfected construct; prey produced from transfected construct.

Overexpression of ex and Mer increases hpo homodimerization.

Source was cell extract of S2 cell line; bait produced from transfected construct; prey produced from endogenous gene.

Either the N- or C-terminus of hpo is sufficient to interact with endogenous hpo.

Source was S2 cell line; bait produced from transfected construct; prey produced from transfected construct.

TEV complementation assay: interaction of bait and prey reconstitutes a functional TEV protease, which in turn cleaves a chimeric fusion of GAL4 and estrogen receptor (ERT2) ligand binding domain. In the absense of TEV activity, the ERT2 ligand binding domain restricts the chimeric protein to the cytosol. Upon cleavage, the free GAL4-VP16 portion of the chimera can activate a UAS-reporter.

Interaction of bait and prey, each attached to complementary fragments of the Venus protein, results in reconstitution of fluorescent activity.

hpo homodimers (as visualized by the reconstituted fluorescent Venus signal) are localized to the apical region of cells in larval wing discs.

Source was live larval wing disc of transgenic fly line; bait produced from tagged transgenic construct; prey produced from tagged transgenic construct.

Source was live S2 cells; bait produced from tagged transgenic construct; prey produced from tagged transgenic construct.

hpo homodimerizes. Mutations that disrupt homodimerization also prevent hpo phosphorylation at residue Tyr195. Mutations that disrupt hpo phosphorylation (T189A and T195A) do not disrupt homodimerization.

Interaction in vitro; protein produced as a recombinant fusion protein in bacterial system.

hpo forms a homodimer in solution.