Source was adult ovary of mutant fly line, heterozygous compared to homozygous; bait produced from endogenous gene; prey produced from endogenous gene.

Treated with RNAse.

Source was adult ovary of mutant fly line, heterozygous compared to homozygous; bait produced from endogenous gene; prey produced from endogenous gene.

Treated with RNAse.

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

In the presence of RNase during immunoprecipitation this interaction was still observed, indicating that it is not mediated through RNA in the RNP complex; may be direct.

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

Treated with RNase.

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

Interaction with this portion of cup protein is not affected by W117A (reported as W106A) mutation in eIF-4E protein.

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

The W117A mutation of the eIF-4E protein does not further reduce the residual interaction observed for the cup Y342A mutation.

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

The Y342A mutation of the cup protein does not further reduce the residual interaction observed for the eIF-4E W117A mutation; the L379A/L383A mutation of cup protein completely eliminates the residual interaction observed for the eIF-4E W117A mutation.

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

In the presence of RNase during immunoprecipitation this interaction was still observed.

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.

Both protein isoforms of eIF-4E tested; no difference observed.

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

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

11 of 24 positives represented clones of eIF-4E.

Two-hybrid system: yeast GAL4-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).

Interaction in vitro; bait produced and labeled by in vitro translation; prey produced and labeled by in vitro translation.

cup and @ compete for binding to eIF-4E@.

Source was cell extract of S2 cell line; bait produced from endogenous gene; prey produced from endogenous gene.

The eIF4E1-cup interaction decreases by treatment with increasing amounts of Thor.

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

Proteins were coexpressed in bacteria to prevent rapid degradation of cup protein. The complex was subjected to limited proteolysis to promote crystallization.

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

Kd = 9.1 nM

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

Mutation of residues on the lateral surface of eIF4E1 (I96A and I112A) specifically disrupts interaction with 4E-BP-type proteins (Thor, cup and 4E-T) but does not affect binding to eIF4G1.

cup was fused to a plasma membrane anchor. The ability of this chimera to recruit prey protein to the plasma membrane was taken as evidence of direct physical interaction.

Source was live S2R+ cells; bait and prey produced from endogenous genes.