Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey produced and labeled by in vitro translation.

The "Region I" and the NLS ("Region II") motifs that mediate binding to cact are similar between Drosophila dl and Dif proteins, but diverge from mammalian Rel proteins.

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

Endogenous Dif activation domain used.

Two-hybrid system: yeast LexA-BD/B42-AD

Positive control.

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

Dif homodimers tested in this assay.

Bait expressed and purified from bacteria.

Prey expressed and purified from bacteria.

cact binds the Rel homology domains of Dif and dl more strongly than that of Rel.

Purified proteins were mixed and precipitated with anti-thioredoxin (Trx tag) antibodies.

Bait expressed and purified from bacteria.

Prey expressed and purified from bacteria.

The cact ankyrin repeat domain binds the Rel homology domains of Dif and dl more strongly than the Rel Rel homology domain.

The interaction of the cact ankyrin repeat domain and the Dif Rel homology domain is not affected by the presence of 200-fold molar excess of the Rel ankyrin repeat domain.