Protein expressed and purified from bacteria.

Electron microscopy shows that purified ttk BTB domain, fused to GST, exists as a tetramer.

Protein expressed and purified from bacteria, then separated from the GST tag by thrombin cleavage.

The isolated ttk–BTB domain elutes from size-exclusion chromatography with an estimated molecular mass of 57.2 kDa, 3.8 times the monomer mass.

Mutation of conserved residues within the ttk BTB domain abolishes the interaction.

Two-hybrid system: yeast LexA/B42.

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

Crosslinked ttk formed mainly high-order multimers.

Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey was previously cloned reagent).

Two-hybrid system: yeast GAL4-BD/GAL4-AD

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

ttk formed mainly high-order multimers.

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

ttk formed mainly high-order multimers.

ttk protein was incubated with biotin labeled DNA containing two copies of its cognate DNA binding motif. DNA was pulled down with streptavidin beads and the amount of luciferase activity pulled down was measured. Unlabeled competitor oligos lacking one or both DNA binding motifs were added in excess to assess the cooperativity of DNA binding by the TF.

Unlabeled competitors containing only one motif were far less effective at competing for binding than oligos containing two nearby motifs. Binding of ttk to its cognate motif is cooperative.

Interaction in vitro; protein produced by in vitro translation.

From an analysis of many TF-ChIP data sets, TF-TF interactions were predicted on the basis of regular DNA sequence motif arrangements. 27 predicted TF-TF interactions were tested using a pull down assay that measures the \'Luminescence Intensity Ratio\'. A cut-off of 7 was used, well above values for six negative controls.

Isoform ttk-PC tested.

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

Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey was previously cloned reagent).

Two-hybrid system: yeast GAL4-BD/GAL4-AD

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

MW values were determined from column calibration with standard proteins. ttk elutes as a multimer.