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/VP16-AD
Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey produced and labeled by in vitro translation.
Mutations in gro that disrupt binding to the WRPW motif typically also disrupt binding of the eh1 motif (FxIxxIL), except for the L692F and L651F mutations, which disrupt only WRPW binding. The V435A and V435L mutations have no effect on gro binding to either the WRPW motif or eh1 domain.
A large scale genetic screen for enhancers of hairy recovered 50 novel gro alleles, including 7 mutations of the gro WD domain. These mutations, along with additional residues identified in a crystal structure of the human homolog TLE, were assessed for their effects on gro interactions with WRPW and eh1 repressor proteins.