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

Together with hb RNA, the nos and pum proteins form a ternary complex.

Three-hybrid system: yeast GAL4-BD/MS2-SL/GAL4-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 GAL4-BD/MS2-SL/GAL4-AD. MS2-SL fused to hb NRE RNA.

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

Recombinant GST-pum was mixed with hb RNA (NRE) and nos, brat and eIF4EHP proteins in various combinations.

Source was adult female ovary of mutant fly line; bait produced from endogenous gene; prey produced from endogenous gene.

The pum-nos interaction is abolished by RNase treatment.

Experiments were done in bam[Δ86] mutant ovaries, which contain only undifferentiated pre-cystoblasts.

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

Ternary complex of nos and pum proteins and hb RNA.

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

nos alters pum RNA-binding specificity. Alone, neither nos nor pum bind to CycB or bcd NRE RNA, but together they form ternary complexes. pum can bind to hb NRE2 RNA alone, but nos increases the strength of the interaction.

Interaction of nos and pum protein binding to RNA, alone or in combination, was tested. hb NRE1, CycB NRE and bcd NRE RNAs were used.

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

nos, pum and Pen mRNA form a ternary complex. Neither pum nor nos bind Pen mRNA on their own in this assay implying a pum-nos interaction.