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

Su(H) binds H specifically. No interaction of Su(H) detected with other nuclear proteins: gro, neur or da.

H inhibits the DNA-binding activity of Su(H).

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

H inhibits the DNA-binding activity of Su(H).

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 LexA-BD/B42-AD

Interaction in vitro; proteins produced by in vitro translation.

H supershifts the Su(H)-DNA complex. H alone cannot bind the DNA probe.

Su(H) and H were assayed for cooperative DNA binding to [32]P labeled 43-mer DNA containing two consensus Su(H) binding sites.

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/B42-AD

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

H causes a supershift in the mobility of the Su(H)-DNA complex. H does not bind this DNA on its own.

H and Su(H) were incubated with [32]P labeled DNA containing a Su(H) binding site.

Prey was cloned reagent (previously isolated).

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

The C-terminal domain of Su(H) is sufficient for binding; addition of N-terminal region, including the first alpha helix, enhances the interaction.

Prey was cloned reagent (previously isolated).

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

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

H and Su(H) form a high affinity 1:1 complex with Kd = 1nM.

Interaction in vitro; proteins produced by in vitro translation.

H supershifts the Su(H)-DNA complex. H alone cannot bind the DNA probe. The intracellular domain of N displaces H to form N-Su(H) DNA-protein complexes. There is no evidence of a N-Su(H)-H trimeric complex formed on the DNA. mam has no noticeable effect on the competition between N-ICN and H for binding to Su(H).

Su(H) and H were assayed for cooperative DNA binding to [32]P labeled DNA probe.

Prey was cloned reagent (previously isolated).

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

Prey was cloned reagent (previously isolated).

Some unexpected combinations exhibit binding: the H NTCT fragment (but not full-length H protein) shows strong binding to the Su(H) CTD[WARE] mutated fragment; the Su(H) CTD fragment (but not full-length Su(H) protein) shows weak binding to H NTCT partial or mutated fragments. The double mutant combinations exhibit no binding.

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

Source was adult heads of transgenic fly line; bait produced from tagged transgenic construct; prey produced from endogenous gene.

Transgenic protein was expressed in the developing eye.

HGScore = 41.465211

Source was cell extract of S2R+ cell line; proteins produced from transfected construct or endogenous gene.

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 LexA-BD/B42-AD

Positive control.

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).

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

H binds Su(H) with Kd = 1nM.

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 lexA-BD/B42-AD

Surface entropy reduction (SER) mutations were introduced into Su(H) to facilitate purification and crystallization: R155T and N281G. These SER mutations do not affect binding.

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

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

Su(H)-H bind with Kd = 2nM.

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

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

Triple Su(H) mutants L434A/L445A/L514A and L445A/L415A/F516A disrupt binding to H.

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 lexA-BD/B42-AD

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 lexA-BD/B42-AD

Source was cell extract of Kc167 cell line; bait produced from transfected construct; prey produced from endogenous gene.

Positive control.

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 lexA-BD/B42-AD

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).

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 lexA-BD/B42-AD

Su(H) and H were mixed with fluorescently labeled DNA. DNA-protein complexes observed by EMSA were used to deduce protein-protein interactions.

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

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).

Myc-trap binding assay; positive control.

Interaction in vitro; bait and prey produced as recombinant fusion proteins in bacterial system.

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 LexA-BD/B42-AD. A previously reported H-LD mutant was used as a negative control.