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/GAL4-AD

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

Source was embryos of wild-type fly line; bait produced from endogenous gene; prey produced from endogenous gene.

Addition of phosphatase does not have an effect on Su(H) interaction with N.

Source was yeast cell line; bait produced as transgenic fusion protein; ancillary factor produced as transgenic protein; prey produced as transgenic fusion protein (prey was produced as transgenic fusion protein.

Three-hybrid system: yeast GAL4-BD/GAL4-AD and an additional ancillary bridging factor.

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

Su(H) binds independently to two different regions of the N intracellular domain: the RAM and PPD domains.

Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey derived from transgenic embryonic extract.

Deletion of both the RAM and PPD domains of N eliminates binding to Su(H).

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

Mutation of both the RAM and PPD domains of N eliminates binding to Su(H).

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

ac did not interact with Su(H) in this experiment.

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

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

Positive control.

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

Positive control.

Interaction in vitro; proteins produced as a recombinant fusion proteins in baculovirus-infected Sf9 insect cell system.

The N-Su(H) complex was calculated to be 170 kDa (close to the 165 kDa predicted).

Interaction in vitro; proteins produced by in vitro translation.

N supershifts the Su(H)-DNA complex. The intracellular domain of N outcompetes H for binding to the Su(H)-DNA complex. 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 N 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.

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.

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

Source was yeast cell line; bait produced as transgenic fusion protein; ancillary factor produced as transgenic protein; prey produced as transgenic fusion protein (prey was produced as transgenic fusion protein.

Three-hybrid system: yeast GAL4-BD/GAL4-AD and an additional ancillary bridging factor.

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

Prey was cloned reagent (previously isolated).

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

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

Positive control.

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

Multiple Su(H) clones identified in this N two hybrid screen.

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

Positive control.

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

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.

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.

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

N binds Su(H) with Kd = 60nM (both the RAM and ANK domains of N are required).

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)-N bind with Kd = 60nM.

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

mam was co-expressed as a bridging factor in this three hybrid experiment, with N as bait and Su(H) as prey.

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

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

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

N intracellular domain aa 1762-2176 was co-expressed as a bridging factor in this three hybrid experiment, with mam as bait and Su(H) as prey.

Kd = 6nM

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

Three-hybrid system: yeast lexA-BD/B42-AD, N ICD was coexpressed as a bridging factor in a yeast three-hybrid assay with mam as bait and Su(H) as prey. Positive control.